{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T13:34:38Z","timestamp":1773063278518,"version":"3.50.1"},"reference-count":64,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,5,18]],"date-time":"2022-05-18T00:00:00Z","timestamp":1652832000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science, Research and Innovation Fund (NSRF)","award":["60169"],"award-info":[{"award-number":["60169"]}]},{"name":"Science Achievement Scholarship of Thailand (SAST), Thailand","award":["60169"],"award-info":[{"award-number":["60169"]}]},{"name":"Post-Doctoral Training Program from the Research Affairs and Graduate School, Khon Kaen University, Thailand","award":["60169"],"award-info":[{"award-number":["60169"]}]},{"name":"Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, Thailand","award":["60169"],"award-info":[{"award-number":["60169"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>There is a desire to develop new molecules that can combat hyperpigmentation. To this end, the N-terminal cysteine-containing heptapeptide TILI-2 has shown promising preliminary results. In this work, the mechanism by which it works was evaluated using a series of biochemical assays focusing on known biochemical pathways, followed by LC-MS\/MS proteomics to discover pathways that have not been considered before. We demonstrate that TILI-2 is a competitive inhibitor of tyrosinase\u2019s monophenolase activity and it could potentially scavenge ABTS and DPPH radicals. It has a very low cytotoxicity up to 1400 \u00b5M against human fibroblast NFDH cells and macrophage-like RAW 264.7 cells. Our proteomics study revealed that another putative mechanism by which TILI-2 may reduce melanin production involves the disruption of the TGF-\u03b2 signaling pathway in mouse B16F1 cells. This result suggests that TILI-2 has potential scope to be used as a depigmenting agent.<\/jats:p>","DOI":"10.3390\/molecules27103228","type":"journal-article","created":{"date-parts":[[2022,5,18]],"date-time":"2022-05-18T23:14:26Z","timestamp":1652915666000},"page":"3228","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Evaluation of TILI-2 as an Anti-Tyrosinase, Anti-Oxidative Agent and Its Role in Preventing Melanogenesis Using a Proteomics Approach"],"prefix":"10.3390","volume":"27","author":[{"given":"Anupong","family":"Joompang","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"},{"name":"Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Preeyanan","family":"Anwised","sequence":"additional","affiliation":[{"name":"Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sompong","family":"Klaynongsruang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"},{"name":"Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3696-8390","authenticated-orcid":false,"given":"Sittiruk","family":"Roytrakul","sequence":"additional","affiliation":[{"name":"Functional Ingredients and Food Innovation Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Khlong Luang, Pathum Thani 12120, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lapatrada","family":"Taemaitree","sequence":"additional","affiliation":[{"name":"Department of Integrated Science, Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2071-2406","authenticated-orcid":false,"given":"Nisachon","family":"Jangpromma","sequence":"additional","affiliation":[{"name":"Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"},{"name":"Department of Integrated Science, Faculty of Science, Khon Kaen University, 123 Mittraphap Road, Muang District, Khon Kaen 40002, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"163","DOI":"10.4103\/2225-4110.113443","article-title":"Evaluation of the antioxidant and melanogenesis inhibitory properties of pracparatum mungo (lu-do huang)","volume":"3","author":"Kao","year":"2013","journal-title":"J. Tradit. Complement. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.jdermsci.2014.07.001","article-title":"Depigmentation caused by application of the active brightening material, rhododendrol, is related to tyrosinase activity at a certain threshold","volume":"76","author":"Kasamatsu","year":"2014","journal-title":"J. Dermatol. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1016\/j.indcrop.2017.10.017","article-title":"Skin-care effects of dandelion leaf extract and stem extract: Antioxidant properties, tyrosinase inhibitory and molecular docking simulations","volume":"111","author":"Xie","year":"2018","journal-title":"Ind. Crops Prod."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"D\u2019Mello, S.A., Finlay, G.J., Baguley, B.C., and Askarian-Amiri, M.E. (2016). Signaling Pathways in Melanogenesis. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17071144"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2080","DOI":"10.1038\/jid.2014.161","article-title":"UV signaling pathways within the skin","volume":"134","author":"Chen","year":"2014","journal-title":"J. Investig. Dermatol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Boo, Y.C. (2020). Up- or Downregulation of Melanin Synthesis Using Amino Acids, Peptides, and Their Analogs. Biomedicines, 8.","DOI":"10.3390\/biomedicines8090322"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Qin, L., Wu, Y., Liu, Y., Chen, Y., and Zhang, P. (2014). Dual effects of alpha-arbutin on monophenolase and diphenolase activities of mushroom tyrosinase. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0109398"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1080\/14756366.2016.1256882","article-title":"Skin whitening agents: Medicinal chemistry perspective of tyrosinase inhibitors","volume":"32","author":"Pillaiyar","year":"2017","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1080\/14756366.2018.1545767","article-title":"A comprehensive review on tyrosinase inhibitors","volume":"34","author":"Zolghadri","year":"2019","journal-title":"J. Enzym. Inhib. Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"781280","DOI":"10.1155\/2012\/781280","article-title":"The Dual Antimelanogenic and Antioxidant Activities of the Essential Oil Extracted from the Leaves of Acorus macrospadiceus (Yamamoto) F. N. Wei et Y. K. Li","volume":"2012","author":"Huang","year":"2012","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1007\/s12272-011-0515-3","article-title":"Inhibition of UVA-mediated melanogenesis by ascorbic acid through modulation of antioxidant defense and nitric oxide system","volume":"34","author":"Panich","year":"2011","journal-title":"Arch. Pharm. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"343","DOI":"10.4062\/biomolther.2013.032","article-title":"Hesperidin Suppresses Melanosome Transport by Blocking the Interaction of Rab27A-Melanophilin","volume":"21","author":"Kim","year":"2013","journal-title":"Biomol. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Upata, M., Siriwoharn, T., Makkhun, S., Yarnpakdee, S., Regenstein, J.M., and Wangtueai, S. (2022). Tyrosinase Inhibitory and Antioxidant Activity of Enzymatic Protein Hydrolysate from Jellyfish (Lobonema smithii). Foods, 11.","DOI":"10.3390\/foods11040615"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1722","DOI":"10.1002\/jsfa.3645","article-title":"Antioxidant and melanogenesis-inhibitory activities of collagen peptide from jellyfish (Rhopilema esculentum)","volume":"89","author":"Zhuang","year":"2009","journal-title":"J. Sci. Food Agric."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1080\/10826068.2020.1789991","article-title":"Extraction, anti-tyrosinase, and antioxidant activities of the collagen hydrolysate derived from Rhopilema hispidum","volume":"51","author":"Salim","year":"2021","journal-title":"Prep. Biochem. Biotechnol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.jbiosc.2015.10.010","article-title":"New tyrosinase inhibitory decapeptide: Molecular insights into the role of tyrosine residues","volume":"121","author":"Ochiai","year":"2016","journal-title":"J. Biosci. Bioeng."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/j.jbiosc.2020.04.002","article-title":"Evaluation of tyrosinase inhibitory activity and mechanism of Leucrocin I and its modified peptides","volume":"130","author":"Joompang","year":"2020","journal-title":"J. Biosci. Bioeng."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2440","DOI":"10.3390\/ijms10062440","article-title":"An updated review of tyrosinase inhibitors","volume":"10","author":"Chang","year":"2009","journal-title":"Int. J. Mol. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6181","DOI":"10.1021\/acs.jafc.5b01026","article-title":"Discovery of Potent Cysteine-Containing Dipeptide Inhibitors against Tyrosinase: A Comprehensive Investigation of 20 x 20 Dipeptides in Inhibiting Dopachrome Formation","volume":"63","author":"Tseng","year":"2015","journal-title":"J. Agric. Food Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3099","DOI":"10.1021\/ci500370x","article-title":"Serendipitous discovery of short peptides from natural products as tyrosinase inhibitors","volume":"54","author":"Hsiao","year":"2014","journal-title":"J. Chem. Inf. Model."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1124\/mol.114.094185","article-title":"Phage display-mediated discovery of novel tyrosinase-targeting tetrapeptide inhibitors reveals the significance of N-terminal preference of cysteine residues and their functional sulfur atom","volume":"87","author":"Lee","year":"2015","journal-title":"Mol. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1016\/j.bbrc.2017.07.158","article-title":"Feruloylserotonin inhibits hydrogen peroxide-induced melanogenesis and apoptosis in B16F10 and SK-Mel-2 melanoma cells","volume":"491","author":"Cho","year":"2017","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1016\/j.biocel.2013.03.006","article-title":"Induction of ATP synthase beta by H2O2 induces melanogenesis by activating PAH and cAMP\/CREB\/MITF signaling in melanoma cells","volume":"45","author":"Kim","year":"2013","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1271\/bbb.62.145","article-title":"Silk protein, sericin, inhibits lipid peroxidation and tyrosinase activity","volume":"62","author":"Kato","year":"1998","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1318","DOI":"10.1016\/j.procbio.2015.05.007","article-title":"A novel peptide purified from the fermented microalga Pavlova lutheri attenuates oxidative stress and melanogenesis in B16F10 melanoma cells","volume":"50","author":"Oh","year":"2015","journal-title":"Process Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e12761","DOI":"10.1111\/jfbc.12761","article-title":"Structural and functional properties of food protein-derived antioxidant peptides","volume":"43","author":"Nwachukwu","year":"2019","journal-title":"J. Food Biochem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"890","DOI":"10.1016\/j.immuni.2017.10.021","article-title":"Granulocyte-Monocyte Progenitors and Monocyte-Dendritic Cell Progenitors Independently Produce Functionally Distinct Monocytes","volume":"47","author":"Yanez","year":"2017","journal-title":"Immunity"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.semcdb.2016.08.006","article-title":"Tissue-specific contribution of macrophages to wound healing","volume":"61","author":"Minutti","year":"2017","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1007\/s12010-009-8526-y","article-title":"Cytotoxicity of bacterial-derived toxins to immortal lung epithelial and macrophage cells","volume":"160","author":"Peterson","year":"2010","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.stem.2009.12.010","article-title":"Key roles for transforming growth factor beta in melanocyte stem cell maintenance","volume":"6","author":"Nishimura","year":"2010","journal-title":"Cell Stem Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1016\/j.molcel.2008.11.002","article-title":"Inhibition of PAX3 by TGF-beta modulates melanocyte viability","volume":"32","author":"Yang","year":"2008","journal-title":"Mol. Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1007\/s11033-008-9304-6","article-title":"Regulation of melanin synthesis by the TGF-beta family in B16 melanoma cells","volume":"36","author":"Murakami","year":"2009","journal-title":"Mol. Biol. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1007\/s10068-019-00668-8","article-title":"Effect of Chrysanthemum indicum Linne extract on melanogenesis through regulation of TGF-beta\/JNK signaling pathway","volume":"28","author":"Hwang","year":"2019","journal-title":"Food Sci. Biotechnol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"750482","DOI":"10.1155\/2009\/750482","article-title":"Role of BMP-4 and Its Signaling Pathways in Cultured Human Melanocytes","volume":"2009","author":"Park","year":"2009","journal-title":"Int. J. Cell Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.jdermsci.2020.06.007","article-title":"TGF-beta3 suppresses melanogenesis in human melanocytes cocultured with UV-irradiated neighboring cells and human skin","volume":"99","author":"Moon","year":"2020","journal-title":"J. Dermatol. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1111\/j.1755-148X.2008.00514.x","article-title":"Pigmentation PAX-ways: The role of Pax3 in melanogenesis, melanocyte stem cell maintenance, and disease","volume":"21","author":"Kubic","year":"2008","journal-title":"Pigment. Cell Melanoma Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/sj.cr.7290105","article-title":"MAPK signal pathways in the regulation of cell proliferation in mammalian cells","volume":"12","author":"Zhang","year":"2002","journal-title":"Cell Res."},{"key":"ref_38","first-page":"153","article-title":"Mitogen-activated protein (MAP) kinase pathways: Regulation and physiological functions","volume":"22","author":"Pearson","year":"2001","journal-title":"Endocr. Rev."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Chae, J.K., Subedi, L., Jeong, M., Park, Y.U., Kim, C.Y., Kim, H., and Kim, S.Y. (2017). Gomisin N Inhibits Melanogenesis through Regulating the PI3K\/Akt and MAPK\/ERK Signaling Pathways in Melanocytes. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18020471"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"9827519","DOI":"10.1155\/2019\/9827519","article-title":"Melanogenic Effects of Maclurin Are Mediated through the Activation of cAMP\/PKA\/CREB and p38 MAPK\/CREB Signaling Pathways","volume":"2019","author":"Hwang","year":"2019","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"14958","DOI":"10.1038\/s41598-018-33352-8","article-title":"Beauvericin inhibits melanogenesis by regulating cAMP\/PKA\/CREB and LXR-alpha\/p38 MAPK-mediated pathways","volume":"8","author":"Lee","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1188","DOI":"10.1080\/09168451.2018.1459176","article-title":"Anti-melanogenic effects of resorcinol are mediated by suppression of cAMP signaling and activation of p38 MAPK signaling","volume":"82","author":"Kang","year":"2018","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1016\/S0092-8674(03)00432-X","article-title":"Mechanisms of TGF-beta signaling from cell membrane to the nucleus","volume":"113","author":"Shi","year":"2003","journal-title":"Cell"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"25307","DOI":"10.1074\/jbc.M600580200","article-title":"Bone morphogenetic protein-4, a novel modulator of melanogenesis","volume":"281","author":"Yaar","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1006\/dbio.2001.0222","article-title":"Wnt and BMP signaling govern lineage segregation of melanocytes in the avian embryo","volume":"233","author":"Jin","year":"2001","journal-title":"Dev. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1111\/ics.12466","article-title":"Intrinsic and extrinsic regulation of human skin melanogenesis and pigmentation","volume":"40","author":"Serre","year":"2018","journal-title":"Int. J. Cosmet. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1530\/rep.1.01075","article-title":"All in the family: TGF-beta family action in testis development","volume":"132","author":"Itman","year":"2006","journal-title":"Reproduction"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1248\/bpb.30.1135","article-title":"Inhibitory effect of p-hydroxybenzyl alcohol on tyrosinase activity and melanogenesis","volume":"30","author":"Liu","year":"2007","journal-title":"Biol. Pharm. Bull."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1661","DOI":"10.3390\/ma5091661","article-title":"Natural Melanogenesis Inhibitors Acting Through the Down-Regulation of Tyrosinase Activity","volume":"5","author":"Chang","year":"2012","journal-title":"Materials"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1002\/ptr.2435","article-title":"Flavonoids, taxifolin and luteolin attenuate cellular melanogenesis despite increasing tyrosinase protein levels","volume":"22","author":"An","year":"2008","journal-title":"Phytother. Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1016\/S0891-5849(98)00315-3","article-title":"Antioxidant activity applying an improved ABTS radical cation decolorization assay","volume":"26","author":"Re","year":"1999","journal-title":"Free Radic. Biol. Med."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1007\/s10930-011-9372-7","article-title":"The effects of temperature and pH on secondary structure and antioxidant activity of Crocodylus siamensis hemoglobin","volume":"31","author":"Jandaruang","year":"2012","journal-title":"Protein J."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1007\/s10930-014-9581-y","article-title":"An investigation of antioxidant and anti-inflammatory activities from blood components of Crocodile (Crocodylus siamensis)","volume":"33","author":"Phosri","year":"2014","journal-title":"Protein J."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Maijaroen, S., Klaynongsruang, S., Roytrakul, S., Konkchaiyaphum, M., Taemaitree, L., and Jangpromma, N. (2022). An Integrated Proteomics and Bioinformatics Analysis of the Anticancer Properties of RT2 Antimicrobial Peptide on Human Colon Cancer (Caco-2) Cells. Molecules, 27.","DOI":"10.3390\/molecules27041426"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/S0021-9258(19)52451-6","article-title":"Protein measurement with the Folin phenol reagent","volume":"193","author":"Lowry","year":"1951","journal-title":"J. Biol. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"174753","DOI":"10.1016\/j.ejphar.2022.174753","article-title":"Proteomic profiling reveals antitumor effects of RT2 peptide on a human colon carcinoma xenograft mouse model","volume":"917","author":"Maijaroen","year":"2022","journal-title":"Eur. J. Pharmacol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1002\/rcm.2898","article-title":"Evaluation of sample fractionation using micro-scale liquid-phase isoelectric focusing on mass spectrometric identification and quantitation of proteins in a SILAC experiment","volume":"21","author":"Thorsell","year":"2007","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"4475","DOI":"10.1002\/pmic.200500921","article-title":"Differential expression analysis of Escherichia coli proteins using a novel software for relative quantitation of LC-MS\/MS data","volume":"6","author":"Johansson","year":"2006","journal-title":"Proteomics"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"3551","DOI":"10.1002\/(SICI)1522-2683(19991201)20:18<3551::AID-ELPS3551>3.0.CO;2-2","article-title":"Probability-based protein identification by searching sequence databases using mass spectrometry data","volume":"20","author":"Perkins","year":"1999","journal-title":"Electrophoresis"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"D183","DOI":"10.1093\/nar\/gkw1138","article-title":"PANTHER version 11: Expanded annotation data from Gene Ontology and Reactome pathways, and data analysis tool enhancements","volume":"45","author":"Mi","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"D380","DOI":"10.1093\/nar\/gkv1277","article-title":"STITCH 5: Augmenting protein-chemical interaction networks with tissue and affinity data","volume":"44","author":"Szklarczyk","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1534","DOI":"10.1292\/jvms.18-0417","article-title":"Identification of reference genes for quantitative PCR analyses in developing mouse gonads","volume":"80","author":"Yokoyama","year":"2018","journal-title":"J. Vet. Med. Sci."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1159\/000228720","article-title":"Identification of suitable normalizing genes for quantitative real-time RT-PCR analysis of gene expression in fetal mouse gonads","volume":"3","author":"Svingen","year":"2009","journal-title":"Sex. Dev."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/10\/3228\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:14:11Z","timestamp":1760138051000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/10\/3228"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,5,18]]},"references-count":64,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2022,5]]}},"alternative-id":["molecules27103228"],"URL":"https:\/\/doi.org\/10.3390\/molecules27103228","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,5,18]]}}}