{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T15:59:38Z","timestamp":1784563178288,"version":"3.55.0"},"reference-count":60,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2023,9,18]],"date-time":"2023-09-18T00:00:00Z","timestamp":1694995200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["Se 460\/21-1"],"award-info":[{"award-number":["Se 460\/21-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["Se 460\/22-1"],"award-info":[{"award-number":["Se 460\/22-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Studies in human colonic cell lines and murine intestine suggest the presence of a Ca2+-activated anion channel, presumably TMEM16a. Is there a potential for fluid secretion in patients with severe cystic fibrosis transmembrane conductance regulator (CFTR) mutations by activating this alternative pathway? Two-dimensional nondifferentiated colonoid\u2013myofibroblast cocultures resembling transit amplifying\/progenitor (TA\/PE) cells, as well as differentiated monolayer (DM) cultures resembling near-surface cells, were established from both healthy controls (HLs) and patients with severe functional defects in the CFTR gene (PwCF). F508del mutant and CFTR knockout (null) mice ileal and colonic mucosa was also studied. HL TA\/PE monolayers displayed a robust short-circuit current response (\u0394Ieq) to UTP (100 \u00b5M), forskolin (Fsk, 10 \u00b5M) and carbachol (CCH, 100 \u00b5M), while \u0394Ieq was much smaller in differentiated monolayers. The selective TMEM16a inhibitor Ani9 (up to 30 \u00b5M) did not alter the response to luminal UTP, significantly decreased Fsk-induced \u0394Ieq, and significantly increased CCH-induced \u0394Ieq in HL TA\/PE colonoid monolayers. The PwCF TA\/PE and the PwCF differentiated monolayers displayed negligible agonist-induced \u0394Ieq, without a significant effect of Ani9. When TMEM16a was localized in intracellular structures, a staining in the apical membrane was not detected. TMEM16a is highly expressed in human colonoid monolayers resembling transit amplifying cells of the colonic cryptal neck zone, from both HL and PwCF. While it may play a role in modulating agonist-induced CFTR-mediated anion currents, it is not localized in the apical membrane, and it has no function as an apical anion channel in cystic fibrosis (CF) and healthy human colonic epithelium.<\/jats:p>","DOI":"10.3390\/ijms241814214","type":"journal-article","created":{"date-parts":[[2023,9,18]],"date-time":"2023-09-18T02:17:30Z","timestamp":1695003450000},"page":"14214","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["The Anion Channel TMEM16a\/Ano1 Modulates CFTR Activity, but Does Not Function as an Apical Anion Channel in Colonic Epithelium from Cystic Fibrosis Patients and Healthy Individuals"],"prefix":"10.3390","volume":"24","author":[{"given":"Azam","family":"Salari","sequence":"first","affiliation":[{"name":"Department of Gastroenterology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renjie","family":"Xiu","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4775-9535","authenticated-orcid":false,"given":"Mahdi","family":"Amiri","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9948-7589","authenticated-orcid":false,"given":"Sophia Theres","family":"Pallenberg","sequence":"additional","affiliation":[{"name":"Department of Pediatric Pneumonology, Allergology and Neonatology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3974-5363","authenticated-orcid":false,"given":"Rainer","family":"Schreiber","sequence":"additional","affiliation":[{"name":"Institute of Physiology, University of Regensburg, 93040 Regensburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9582-1025","authenticated-orcid":false,"given":"Anna-Maria","family":"Dittrich","sequence":"additional","affiliation":[{"name":"Department of Pediatric Pneumonology, Allergology and Neonatology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Burkhard","family":"T\u00fcmmler","sequence":"additional","affiliation":[{"name":"Department of Pediatric Pneumonology, Allergology and Neonatology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6222-4593","authenticated-orcid":false,"given":"Karl","family":"Kunzelmann","sequence":"additional","affiliation":[{"name":"Institute of Physiology, University of Regensburg, 93040 Regensburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9600-2769","authenticated-orcid":false,"given":"Ursula","family":"Seidler","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1056","DOI":"10.1172\/JCI73193","article-title":"Defective goblet cell exocytosis contributes to murine cystic fibrosis-associated intestinal disease","volume":"125","author":"Liu","year":"2015","journal-title":"J. Clin. Investig."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2613","DOI":"10.1172\/JCI38662","article-title":"Normal mouse intestinal mucus release requires cystic fibrosis transmembrane regulator-dependent bicarbonate secretion","volume":"119","author":"Garcia","year":"2009","journal-title":"J. Clin. Investig."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1084\/jem.20120562","article-title":"Bicarbonate and functional CFTR channel are required for proper mucin secretion and link cystic fibrosis with its mucus phenotype","volume":"209","author":"Gustafsson","year":"2012","journal-title":"J. Exp. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/S0140-6736(08)61162-9","article-title":"Cystic fibrosis: Impaired bicarbonate secretion and mucoviscidosis","volume":"372","author":"Quinton","year":"2008","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1016\/j.jcf.2010.06.003","article-title":"Quantitative expression analyses of candidates for alternative anion conductance in cystic fibrosis mouse models","volume":"9","author":"Braun","year":"2010","journal-title":"J. Cyst. Fibros. Off. J. Eur. Cyst. Fibros. Soc."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"693","DOI":"10.2147\/JEP.S255377","article-title":"Pharmacological Modulation of Ion Channels for the Treatment of Cystic Fibrosis","volume":"13","author":"Pinto","year":"2021","journal-title":"J. Exp. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.coph.2017.10.002","article-title":"Bypassing CFTR dysfunction in cystic fibrosis with alternative pathways for anion transport","volume":"34","author":"Li","year":"2017","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"G617","DOI":"10.1152\/ajpgi.2000.278.4.G617","article-title":"Defective cholinergic Cl\u2212 secretion and detection of K+ secretion in rectal biopsies from cystic fibrosis patients","volume":"278","author":"Mall","year":"2000","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1262","DOI":"10.3389\/fphys.2019.01262","article-title":"Control of Ion Transport by Tmem16a Expressed in Murine Intestine","volume":"10","author":"Kunzelmann","year":"2019","journal-title":"Front. Physiol."},{"key":"ref_10","first-page":"461","article-title":"Determinants of mild clinical symptoms in cystic fibrosis patients","volume":"93","author":"Veeze","year":"1994","journal-title":"Residual chloride secretion measured in rectal biopsies in relation to the genotype. J. Clin. Investig."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1038\/ng0396-280","article-title":"Modulation of disease severity in cystic fibrosis transmembrane conductance regulator deficient mice by a secondary genetic factor","volume":"12","author":"Rozmahel","year":"1996","journal-title":"Nat. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1006\/bbrc.1996.0306","article-title":"In vivo measurements of ion transport in long-living CF mice","volume":"219","author":"Wilschanski","year":"1996","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s12276-019-0299-y","article-title":"Anoctamin 1\/TMEM16A controls intestinal Cl\u2212 secretion induced by carbachol and cholera toxin","volume":"51","author":"Lee","year":"2019","journal-title":"Exp. Mol. Med."},{"key":"ref_14","first-page":"100912","article-title":"Intestinal TMEM16A control luminal chloride secretion in a NHERF1 dependent manner","volume":"25","author":"Saha","year":"2021","journal-title":"Biochem. Biophys. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5317","DOI":"10.1113\/jphysiol.2012.232124","article-title":"Rescue of epithelial HCO3- secretion in murine intestine by apical membrane expression of the cystic fibrosis transmembrane conductance regulator mutant F508del","volume":"590","author":"Xiao","year":"2012","journal-title":"J. Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1038\/nm.3201","article-title":"A functional CFTR assay using primary cystic fibrosis intestinal organoids","volume":"19","author":"Dekkers","year":"2013","journal-title":"Nat. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"638","DOI":"10.1053\/j.gastro.2015.11.047","article-title":"Human Enteroids as a Model of Upper Small Intestinal Ion Transport Physiology and Pathophysiology","volume":"150","author":"In","year":"2016","journal-title":"Gastroenterology"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Salari, A., Zhou, K., Nikolovska, K., Seidler, U., and Amiri, M. (2023). Human Colonoid-Myofibroblast Coculture for Study of Apical Na+\/H+ Exchangers of the Lower Cryptal Neck Region. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24054266"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1172\/JCI116966","article-title":"Localization of cystic fibrosis transmembrane conductance regulator mRNA in the human gastrointestinal tract by in situ hybridization","volume":"93","author":"Strong","year":"1994","journal-title":"J. Clin. Investig."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"694","DOI":"10.3389\/fphys.2019.00694","article-title":"Normal Calcium-Activated Anion Secretion in a Mouse Selectively Lacking TMEM16A in Intestinal Epithelium","volume":"10","author":"Vega","year":"2019","journal-title":"Front. Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"G1370","DOI":"10.1152\/ajpgi.00074.2009","article-title":"Ano1 is a selective marker of interstitial cells of Cajal in the human and mouse gastrointestinal tract","volume":"296","author":"Gibbons","year":"2009","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Centeio, R., Cabrita, I., Benedetto, R., Talbi, K., Ousingsawat, J., Schreiber, R., Sullivan, J.K., and Kunzelmann, K. (2020). Pharmacological Inhibition and Activation of the Ca2+ Activated Cl\u2212 Channel TMEM16A. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21072557"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Seo, Y., Lee, H.K., Park, J., Jeon, D.K., Jo, S., Jo, M., and Namkung, W. (2016). Ani9, A Novel Potent Small-Molecule ANO1 Inhibitor with Negligible Effect on ANO2. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0155771"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"12990","DOI":"10.1074\/jbc.M109.068544","article-title":"Tmem16A encodes the Ca2+-activated Cl- channel in mouse submandibular salivary gland acinar cells","volume":"285","author":"Romanenko","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6141","DOI":"10.1113\/jphysiol.2012.240838","article-title":"Association of TMEM16A chloride channel overexpression with airway goblet cell metaplasia","volume":"590","author":"Scudieri","year":"2012","journal-title":"J. Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ousingsawat, J., Centeio, R., Schreiber, R., and Kunzelmann, K. (2022). Expression of SLC26A9 in Airways and Its Potential Role in Asthma. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23062998"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1007\/s00424-007-0248-9","article-title":"Distal colonic Na+ absorption inhibited by luminal P2Y2 receptors","volume":"454","author":"Matos","year":"2007","journal-title":"Pflug. Arch. Eur. J. Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1038\/sj.bjp.0706353","article-title":"The role of epithelial P2Y2 and P2Y4 receptors in the regulation of intestinal chloride secretion","volume":"146","author":"Ghanem","year":"2005","journal-title":"Br. J. Pharmacol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"C1862","DOI":"10.1152\/ajpcell.1997.272.6.C1862","article-title":"Role of purinergic receptors in chloride secretion in Caco-2 cells","volume":"272","author":"Inoue","year":"1997","journal-title":"Am. J. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1152\/physrev.00039.2011","article-title":"Structure and function of TMEM16 proteins (anoctamins)","volume":"94","author":"Pedemonte","year":"2014","journal-title":"Physiol. Rev."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Danahay, H., and Gosling, M. (2020). TMEM16A: An Alternative Approach to Restoring Airway Anion Secretion in Cystic Fibrosis?. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21072386"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1080\/17476348.2023.2228194","article-title":"Thinking outside the box: A review of gastrointestinal symptoms and complications in cystic fibrosis","volume":"17","author":"Yule","year":"2023","journal-title":"Expert Rev. Respir. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1931","DOI":"10.1002\/ppul.26413","article-title":"Critical disease burdens of Australian adults with cystic fibrosis: Results from an online survey","volume":"58","author":"Ward","year":"2023","journal-title":"Pediatr. Pulmonol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/j.jcf.2022.10.006","article-title":"Multicenter prospective study showing a high gastrointestinal symptom burden in cystic fibrosis","volume":"22","author":"Moshiree","year":"2023","journal-title":"J. Cyst. Fibros. Off. J. Eur. Cyst. Fibros. Soc."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"101788","DOI":"10.1016\/j.bpg.2022.101788","article-title":"Digestive outcomes in Cystic fibrosis","volume":"56\u201357","author":"Ley","year":"2022","journal-title":"Best. Pract. Res. Clin. Gastroenterol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Rodenburg, L.W., Delpiano, L., Railean, V., Centeio, R., Pinto, M.C., Smits, S.M.A., van der Windt, I.S., van Hugten, C.F.J., van Beuningen, S.F.B., and Rodenburg, R.N.P. (2022). Drug Repurposing for Cystic Fibrosis: Identification of Drugs That Induce CFTR-Independent Fluid Secretion in Nasal Organoids. Int. J. Mol. Sci., 23.","DOI":"10.1101\/2022.09.23.509034"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1018","DOI":"10.1111\/bph.15323","article-title":"Inhibition of Na+\/H+ exchanger isoform 3 improves gut fluidity and alkalinity in cystic fibrosis transmembrane conductance regulator-deficient and F508del mutant mice","volume":"178","author":"Tan","year":"2021","journal-title":"Br. J. Pharmacol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Tan, X.J., Kini, A., Romermann, D., and Seidler, U. (2022). The NHE3 Inhibitor Tenapanor Prevents Intestinal Obstructions in CFTR-Deleted Mice. Int. J. Mol. Sci., 23.","DOI":"10.22541\/au.165348030.02708198\/v1"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1763","DOI":"10.1113\/jphysiol.2009.181933","article-title":"Adrenaline-induced colonic K+ secretion is mediated by KCa1.1 (BK) channels","volume":"588","author":"Sausbier","year":"2010","journal-title":"J. Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1111\/j.1469-7793.1997.411bb.x","article-title":"A functional CFTR protein is required for mouse intestinal cAMP-, cGMP- and Ca2+-dependent HCO3\u2212 secretion","volume":"505","author":"Seidler","year":"1997","journal-title":"J. Physiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1164\/rccm.202201-0219OC","article-title":"Effects of Elexacaftor\/Tezacaftor\/Ivacaftor Therapy on Lung Clearance Index and Magnetic Resonance Imaging in Patients with Cystic Fibrosis and One or Two F508del Alleles","volume":"206","author":"Graeber","year":"2022","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_42","first-page":"4502","article-title":"TMEM16A is indispensable for basal mucus secretion in airways and intestine","volume":"33","author":"Benedetto","year":"2019","journal-title":"FASEB J. Off. Publ. Fed. Am. Soc. Exp. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.jcmgh.2017.11.013","article-title":"Cftr Modulates Wnt\/beta-Catenin Signaling and Stem Cell Proliferation in Murine Intestine","volume":"5","author":"Strubberg","year":"2018","journal-title":"Cell. Mol. Gastroenterol. Hepatol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e13774","DOI":"10.1111\/apha.13774","article-title":"Sodium\/hydrogen-exchanger-2 modulates colonocyte lineage differentiation","volume":"234","author":"Nikolovska","year":"2022","journal-title":"Acta Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"590262","DOI":"10.3389\/fphys.2020.590262","article-title":"Calcium-Activated Chloride Channel ANO1\/TMEM16A: Regulation of Expression and Signaling","volume":"11","author":"Dulin","year":"2020","journal-title":"Front. Physiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.bbadis.2006.01.007","article-title":"Ex vivo biochemical analysis of CFTR in human rectal biopsies","volume":"1762","author":"Stanke","year":"2006","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_47","first-page":"e59815","article-title":"Real-Time, Semi-Automated Fluorescent Measurement of the Airway Surface Liquid pH of Primary Human Airway Epithelial Cells","volume":"9","author":"Haq","year":"2019","journal-title":"J. Vis. Exp."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/j.resp.2007.06.005","article-title":"Liquid movement across the surface epithelium of large airways","volume":"159","author":"Chambers","year":"2007","journal-title":"Respir. Physiol. Neurobiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1136\/jmg.2007.053561","article-title":"Diversity of the basic defect of homozygous CFTR mutation genotypes in humans","volume":"45","author":"Stanke","year":"2008","journal-title":"J. Med. Genet."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1164\/rccm.202008-3176LE","article-title":"Safety and Efficacy of Elexacaftor\/Tezacaftor\/Ivacaftor for 24 Weeks or Longer in People with Cystic Fibrosis and One or More F508del Alleles: Interim Results of an Open-Label Phase 3 Clinical Trial","volume":"203","author":"Griese","year":"2021","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1172\/JCI118917","article-title":"A Delta F508 mutation in mouse cystic fibrosis transmembrane conductance regulator results in a temperature-sensitive processing defect in vivo","volume":"98","author":"French","year":"1996","journal-title":"J. Clin. Investig."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1002\/ibd.21744","article-title":"Loss of downregulated in adenoma (DRA) impairs mucosal HCO3-secretion in murine ileocolonic inflammation","volume":"18","author":"Xiao","year":"2012","journal-title":"Inflamm. Bowel Dis."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1762","DOI":"10.1053\/j.gastro.2011.07.050","article-title":"Long-term Expansion of Epithelial Organoids From Human Colon, Adenoma, Adenocarcinoma, and Barrett's Epithelium","volume":"141","author":"Sato","year":"2011","journal-title":"Gastroenterology"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Khalil, H., Nie, W.X., Edwards, R.A., and Yoo, J. (2013). Isolation of Primary Myofibroblasts from Mouse and Human Colon Tissue. J. Vis. Exp., e50611.","DOI":"10.3791\/50611-v"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1756","DOI":"10.1038\/s41596-019-0160-8","article-title":"High-resolution 3D imaging of fixed and cleared organoids","volume":"14","author":"Dekkers","year":"2019","journal-title":"Nat. Protoc."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1016\/j.jmb.2006.10.086","article-title":"Domain interdependence in the biosynthetic assembly of CFTR","volume":"365","author":"Cui","year":"2007","journal-title":"J. Mol. Biol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"e67292","DOI":"10.7554\/eLife.67292","article-title":"Intestinal goblet cells sample and deliver lumenal antigens by regulated endocytic uptake and transcytosis","volume":"10","author":"Gustafsson","year":"2021","journal-title":"eLife"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.bcp.2020.114040","article-title":"Deciphering ion transporters, kinases and PDZ-adaptor molecules that mediate guanylate cyclase C agonist-dependent intestinal fluid loss in vivo","volume":"178","author":"Liu","year":"2020","journal-title":"Biochem. Pharmacol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"W289","DOI":"10.1093\/nar\/gki390","article-title":"PRALINE: A multiple sequence alignment toolbox that integrates homology-extended and secondary structure information","volume":"33","author":"Simossis","year":"2005","journal-title":"Nucleic Acids Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/14214\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:52:42Z","timestamp":1760129562000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/14214"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,18]]},"references-count":60,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2023,9]]}},"alternative-id":["ijms241814214"],"URL":"https:\/\/doi.org\/10.3390\/ijms241814214","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,18]]}}}