{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T16:58:10Z","timestamp":1790873890085,"version":"4.1.0"},"reference-count":61,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T00:00:00Z","timestamp":1745539200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Chronic oxidative distress results in cellular damage, necessitating adaptive mechanisms for redox balance. The transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) is pivotal in the regulation of key antioxidant and cytoprotective genes. Under normal conditions, Nrf2 undergoes rapid degradation through polyubiquitination. However, it can be activated during oxidative eustress and distress via modifications of its inhibitor Kelch-like ECH-associated protein 1 (KEAP1). Activation of the Nrf2-Keap1 signaling pathway may decelerate aging-related muscle degeneration, such as sarcopenia and cachexia. In this study, we investigated the efficacy of two muscle-active endogenous factors, creatine and L-\u03b2-aminoisobutyric acid (L-BAIBA), as well as two natural phytochemicals, luteolin and silibinin, to induce Nrf2 in the murine myoblast cell line C2C12. Our results revealed that only luteolin significantly enhances Nrf2 activity in both proliferating and differentiated C2C12 cells, leading to increased expression of Nrf2 target genes in proliferating C2C12 cells. In contrast, the other three compounds had either no or only minor effects on Nrf2 activity or target gene expression. Our results underscore the distinct responses of C2C12 cells to different Nrf2 activators, emphasizing the significance of cellular context in their biological effects and highlight luteolin as a potential future treatment option to counteract muscle wasting associated with sarcopenia and cachexia.<\/jats:p>","DOI":"10.3390\/ijms26094092","type":"journal-article","created":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T08:02:57Z","timestamp":1745568177000},"page":"4092","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Luteolin Induces Nrf2 Activity in C2C12 Cells: Implications for Muscle Health"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-9030-9329","authenticated-orcid":false,"given":"Nicole","family":"B\u00f6ttcher","sequence":"first","affiliation":[{"name":"Department of Anatomy and Cell Biology, Uniklinik RWTH Aachen, 52074 Aachen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frank","family":"Suhr","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, Uniklinik RWTH Aachen, 52074 Aachen, Germany"},{"name":"Division of Molecular Exercise Physiology, Faculty of Life Sciences: Food, Nutrition and Health, University of Bayreuth, 95326 Kulmbach, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7886-9211","authenticated-orcid":false,"given":"Thomas","family":"Pufe","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, Uniklinik RWTH Aachen, 52074 Aachen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christoph Jan","family":"Wruck","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, Uniklinik RWTH Aachen, 52074 Aachen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6973-3080","authenticated-orcid":false,"given":"Athanassios","family":"Fragoulis","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, Uniklinik RWTH Aachen, 52074 Aachen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1169","DOI":"10.1152\/physrev.00023.2017","article-title":"The KEAP1-NRF2 System: A Thiol-Based Sensor-Effector Apparatus for Maintaining Redox Homeostasis","volume":"98","author":"Yamamoto","year":"2018","journal-title":"Physiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3486","DOI":"10.1128\/MCB.00180-12","article-title":"Structural and functional characterization of Nrf2 degradation by the glycogen synthase kinase 3\/beta-TrCP axis","volume":"32","author":"Rada","year":"2012","journal-title":"Mol. Cell. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1655","DOI":"10.1128\/MCB.01095-15","article-title":"Nrf2-Mediated Regulation of Skeletal Muscle Glycogen Metabolism","volume":"36","author":"Uruno","year":"2016","journal-title":"Mol. Cell. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1007\/s00421-013-2806-5","article-title":"Exercise-induced oxidative stress and hypoxic exercise recovery","volume":"114","author":"Ballmann","year":"2014","journal-title":"Eur. J. Appl. Physiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.freeradbiomed.2014.02.004","article-title":"Lifelong training preserves some redox-regulated adaptive responses after an acute exercise stimulus in aged human skeletal muscle","volume":"70","author":"Cobley","year":"2014","journal-title":"Free Radic. Biol. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1487","DOI":"10.1016\/j.freeradbiomed.2010.08.010","article-title":"Dysfunctional Nrf2-Keap1 redox signaling in skeletal muscle of the sedentary old","volume":"49","author":"Safdar","year":"2010","journal-title":"Free Radic. Biol. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1852","DOI":"10.1091\/mbc.e12-09-0666","article-title":"Inhibition of nuclear factor-erythroid 2-related factor (Nrf2) by caveolin-1 promotes stress-induced premature senescence","volume":"24","author":"Volonte","year":"2013","journal-title":"Mol. Biol. Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.freeradbiomed.2016.04.024","article-title":"Exercise-induced Nrf2-signaling is impaired in aging","volume":"96","author":"Done","year":"2016","journal-title":"Free Radic. Biol. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4215","DOI":"10.1113\/JP286460","article-title":"Muscle-derived IL-1beta regulates EcSOD expression via the NBR1-p62-Nrf2 pathway in muscle during cancer cachexia","volume":"602","author":"Yamada","year":"2024","journal-title":"J. Physiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2636","DOI":"10.1016\/S0140-6736(19)31138-9","article-title":"Sarcopenia","volume":"393","author":"Sayer","year":"2019","journal-title":"Lancet"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1093\/ageing\/afq034","article-title":"Sarcopenia: European consensus on definition and diagnosis: Report of the European Working Group on Sarcopenia in Older People","volume":"39","author":"Baeyens","year":"2010","journal-title":"Age Ageing"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1-1","DOI":"10.1096\/fasebj.2020.34.s1.05173","article-title":"Muscle-specific Nrf2 deficiency exacerbates cancer cachexia-induced skeletal muscle atrophy","volume":"34","author":"Okutsu","year":"2020","journal-title":"FASEB J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1128\/MCB.01080-08","article-title":"The antioxidant defense system Keap1-Nrf2 comprises a multiple sensing mechanism for responding to a wide range of chemical compounds","volume":"29","author":"Kobayashi","year":"2009","journal-title":"Mol. Cell. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4011","DOI":"10.1016\/j.bmc.2013.04.019","article-title":"Small molecules inhibit the interaction of Nrf2 and the Keap1 Kelch domain through a non-covalent mechanism","volume":"21","author":"Marcotte","year":"2013","journal-title":"Bioorg. Med. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2035","DOI":"10.2165\/00003495-200161140-00003","article-title":"The use of silymarin in the treatment of liver diseases","volume":"61","author":"Saller","year":"2001","journal-title":"Drugs"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"108393","DOI":"10.1016\/j.jmgm.2022.108393","article-title":"Structural basis of Nrf2 activation by flavonolignans from silymarin","volume":"119","author":"Bello","year":"2023","journal-title":"J. Mol. Graph. Model."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Vargas-Mendoza, N., Morales-Gonzalez, A., Morales-Martinez, M., Soriano-Ursua, M.A., Delgado-Olivares, L., Sandoval-Gallegos, E.M., Madrigal-Bujaidar, E., Alvarez-Gonzalez, I., Madrigal-Santillan, E., and Morales-Gonzalez, J.A. (2020). Flavolignans from Silymarin as Nrf2 Bioactivators and Their Therapeutic Applications. Biomedicines, 8.","DOI":"10.3390\/biomedicines8050122"},{"key":"ref_18","unstructured":"Li, Y., Li, H., Sun, M., Chen, H., Xiao, Y., Wang, J., Zhang, Y., Fang, S., and Kou, J. Silibinin alleviates acute liver failure by modulating AKT\/GSK3beta\/Nrf2\/GPX4 pathway, Naunyn-Schmiedebergs Arch. Pharmacol."},{"key":"ref_19","first-page":"57","article-title":"Luteolin protects rat PC12 and C6 cells against MPP+ induced toxicity via an ERK dependent Keap1-Nrf2-ARE pathway","volume":"72","author":"Wruck","year":"2007","journal-title":"J. Neural Transm. Suppl."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Kitakaze, T., Makiyama, A., Yamashita, Y., and Ashida, H. (2020). Low dose of luteolin activates Nrf2 in the liver of mice at start of the active phase but not that of the inactive phase. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0231403"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3159358","DOI":"10.1155\/2017\/3159358","article-title":"Antifatigue Effect of Luteolin-6-C-Neohesperidoside on Oxidative Stress Injury Induced by Forced Swimming of Rats through Modulation of Nrf2\/ARE Signaling Pathways","volume":"2017","author":"Duan","year":"2017","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.abb.2019.01.012","article-title":"A physiological concentration of luteolin induces phase II drug-metabolizing enzymes through the ERK1\/2 signaling pathway in HepG2 cells","volume":"663","author":"Kitakaze","year":"2019","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1186\/s10020-023-00729-z","article-title":"beta-aminoisobutyrics acid, a metabolite of BCAA, activates the AMPK\/Nrf-2 pathway to prevent ferroptosis and ameliorates lung ischemia-reperfusion injury","volume":"29","author":"Zhang","year":"2023","journal-title":"Mol. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1931","DOI":"10.1128\/MCB.00118-16","article-title":"AMPK Facilitates Nuclear Accumulation of Nrf2 by Phosphorylating at Serine 550","volume":"36","author":"Joo","year":"2016","journal-title":"Mol. Cell. Biol."},{"key":"ref_25","first-page":"1009","article-title":"Effects of different-intensity exercise and creatine supplementation on mitochondrial biogenesis and redox status in mice","volume":"25","author":"Taskin","year":"2022","journal-title":"Iran. J. Basic Med. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Leem, Y.H., Park, J.S., Park, J.E., Kim, D.Y., and Kim, H.S. (2024). Creatine supplementation with exercise reduces alpha-synuclein oligomerization and necroptosis in Parkinson\u2019s disease mouse model. J. Nutr. Biochem., 126.","DOI":"10.1016\/j.jnutbio.2024.109586"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1599","DOI":"10.1016\/j.freeradbiomed.2011.03.008","article-title":"Luteolin inhibits Nrf2 leading to negative regulation of the Nrf2\/ARE pathway and sensitization of human lung carcinoma A549 cells to therapeutic drugs","volume":"50","author":"Tang","year":"2011","journal-title":"Free Radic. Biol. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1038\/270725a0","article-title":"Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle","volume":"270","author":"Yaffe","year":"1977","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Ryter, S.W. (2022). Heme Oxygenase-1: An Anti-Inflammatory Effector in Cardiovascular, Lung, and Related Metabolic Disorders. Antioxidants, 11.","DOI":"10.3390\/antiox11030555"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ross, D., and Siegel, D. (2021). The diverse functionality of NQO1 and its roles in redox control. Redox Biol., 41.","DOI":"10.1016\/j.redox.2021.101950"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1376","DOI":"10.1096\/fj.09-146308","article-title":"Increased superoxide in vivo accelerates age-associated muscle atrophy through mitochondrial dysfunction and neuromuscular junction degeneration","volume":"24","author":"Jang","year":"2010","journal-title":"FASEB J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1007\/s12603-018-1139-9","article-title":"International Clinical Practice Guidelines for Sarcopenia (ICFSR): Screening, Diagnosis and Management","volume":"22","author":"Dent","year":"2018","journal-title":"J. Nutr. Health Aging"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1093\/gerona\/glaa309","article-title":"Omega-3 Supplementation Improves Isometric Strength But Not Muscle Anabolic and Catabolic Signaling in Response to Resistance Exercise in Healthy Older Adults","volume":"76","author":"Dalle","year":"2021","journal-title":"J. Gerontol. Ser. A"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.jphys.2014.06.012","article-title":"Adherence to exercise programs for older people is influenced by program characteristics and personal factors: A systematic review","volume":"60","author":"Picorelli","year":"2014","journal-title":"J. Physiother."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"20428","DOI":"10.18632\/oncotarget.14670","article-title":"Skeletal muscle aging: Influence of oxidative stress and physical exercise","volume":"8","author":"Gomes","year":"2017","journal-title":"Oncotarget"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1016\/0005-2728(70)90220-3","article-title":"One-electron-transfer reactions in biochemical systems. V. Difference in the mechanism of quinone reduction by the NADH dehydrogenase and the NAD(P)H dehydrogenase (DT-diaphorase)","volume":"216","author":"Iyanagi","year":"1970","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"13593","DOI":"10.1038\/ncomms13593","article-title":"NQO1 inhibits proteasome-mediated degradation of HIF-1alpha","volume":"7","author":"Oh","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1101\/gad.319905","article-title":"A mechanism of ubiquitin-independent proteasomal degradation of the tumor suppressors p53 and p73","volume":"19","author":"Asher","year":"2005","journal-title":"Genes Dev."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Luo, J.F., Shen, X.Y., Lio, C.K., Dai, Y., Cheng, C.S., Liu, J.X., Yao, Y.D., Yu, Y., Xie, Y., and Luo, P. (2018). Activation of Nrf2\/HO-1 Pathway by Nardochinoid C Inhibits Inflammation and Oxidative Stress in Lipopolysaccharide-Stimulated Macrophages. Front. Pharmacol., 9.","DOI":"10.3389\/fphar.2018.00911"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"7840","DOI":"10.4049\/jimmunol.179.11.7840","article-title":"Heme oxygenase-1 inhibits the expression of adhesion molecules associated with endothelial cell activation via inhibition of NF-kappaB RelA phosphorylation at serine 276","volume":"179","author":"Seldon","year":"2007","journal-title":"J. Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"11624","DOI":"10.1038\/ncomms11624","article-title":"Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription","volume":"7","author":"Kobayashi","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"117823","DOI":"10.1016\/j.lfs.2020.117823","article-title":"Sulforaphane protects against skeletal muscle dysfunction in spontaneous type 2 diabetic db\/db mice","volume":"255","author":"Wang","year":"2020","journal-title":"Life Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.jinorgbio.2017.11.010","article-title":"Dietary luteolin protects against HgCl(2)-induced renal injury via activation of Nrf2-mediated signaling in rat","volume":"179","author":"Tan","year":"2018","journal-title":"J. Inorg. Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Tan, X., Yang, Y., Xu, J., Zhang, P., Deng, R., Mao, Y., He, J., Chen, Y., Zhang, Y., and Ding, J. (2019). Luteolin Exerts Neuroprotection via Modulation of the p62\/Keap1\/Nrf2 Pathway in Intracerebral Hemorrhage. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.01551"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Rajput, S.A., Shaukat, A., Wu, K., Rajput, I.R., Baloch, D.M., Akhtar, R.W., Raza, M.A., Najda, A., Rafal, P., and Albrakati, A. (2021). Luteolin Alleviates AflatoxinB(1)-Induced Apoptosis and Oxidative Stress in the Liver of Mice through Activation of Nrf2 Signaling Pathway. Antioxidants, 10.","DOI":"10.3390\/antiox10081268"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Luo, C., Huang, P., Cheng, Y., Ma, Y., Gao, J., and Ding, H. (2025). Luteolin alleviates muscle atrophy, mitochondrial dysfunction and abnormal FNDC5 expression in high fat diet-induced obese rats and palmitic acid-treated C2C12 myotubes. J. Nutr. Biochem., 135.","DOI":"10.1016\/j.jnutbio.2024.109780"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1016\/j.bbrc.2014.04.039","article-title":"Luteolin inhibits the Nrf2 signaling pathway and tumor growth in vivo","volume":"447","author":"Chian","year":"2014","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"105526","DOI":"10.1016\/j.phrs.2021.105526","article-title":"The hormetic dose-response mechanism: Nrf2 activation","volume":"167","author":"Calabrese","year":"2021","journal-title":"Pharmacol. Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1080\/01480545.2018.1504961","article-title":"Luteolin protects microglia against rotenone-induced toxicity in a hormetic manner through targeting oxidative stress response, genes associated with Parkinson\u2019s disease and inflammatory pathways","volume":"43","author":"Elmazoglu","year":"2020","journal-title":"Drug Chem. Toxicol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1021\/jf020926l","article-title":"Simultaneous determination of all polyphenols in vegetables, fruits, and teas","volume":"51","author":"Sakakibara","year":"2003","journal-title":"J. Agric. Food Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1016\/j.foodchem.2007.04.038","article-title":"Antioxidant activity and phenolic compounds in 32 selected herbs","volume":"105","author":"Czemerys","year":"2007","journal-title":"Food Chem."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Casili, G., Ardizzone, A., Lanza, M., Gugliandolo, E., Portelli, M., Militi, A., Cuzzocrea, S., Esposito, E., and Paterniti, I. (2020). Treatment with Luteolin Improves Lipopolysaccharide-Induced Periodontal Diseases in Rats. Biomedicines, 8.","DOI":"10.3390\/biomedicines8100442"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1146\/annurev.nutr.22.111401.144957","article-title":"Dietary flavonoids: Bioavailability, metabolic effects, and safety","volume":"22","author":"Ross","year":"2002","journal-title":"Annu. Rev. Nutr."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1002\/biof.1689","article-title":"Luteolin supplements: All that glitters is not gold","volume":"47","author":"Theoharides","year":"2021","journal-title":"Biofactors"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Tie, F., Fu, Y., Hu, N., and Wang, H. (2022). Silibinin Protects against H2O2-Induced Oxidative Damage in SH-SY5Y Cells by Improving Mitochondrial Function. Antioxidants, 11.","DOI":"10.3390\/antiox11061101"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"5694223","DOI":"10.1155\/2022\/5694223","article-title":"Silibinin Alleviates Muscle Atrophy Caused by Oxidative Stress Induced by Cisplatin through ERK\/FoxO and JNK\/FoxO Pathways","volume":"2022","author":"Chi","year":"2022","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1111\/jnc.13959","article-title":"Psoriasin has divergent effects on the innate immune responses of murine glial cells","volume":"141","author":"Jansen","year":"2017","journal-title":"J. Neurochem."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/S0304-3940(02)01423-4","article-title":"Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data","volume":"339","author":"Ramakers","year":"2003","journal-title":"Neurosci. Lett."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Fragoulis, A., Biller, K., Fragoulis, S., Lex, D., Uhlig, S., and Reiss, L.K. (2021). Reference Gene Selection for Gene Expression Analyses in Mouse Models of Acute Lung Injury. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22157853"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/9\/4092\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:21:35Z","timestamp":1760030495000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/9\/4092"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,25]]},"references-count":61,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,5]]}},"alternative-id":["ijms26094092"],"URL":"https:\/\/doi.org\/10.3390\/ijms26094092","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,25]]}}}