Fig. 5

PCDH20 interacted with downstream ATF6. a Enriched Ontology clusters heatmap of predicted PCDH20 interactors. b Immunoblot of ATF6 expression in Caco-2 cell line with stable PCDH20 knockdown and overexpression. c Immunoblot of ATF6 expression in PCDH20-deficient Caco-2 cell line transfected with PCDH20 overexpression plasmid. d Decreased ATF6 expression in PCDH20-deficient CD patients using immunofluorescence. Magnification × 200, scale bars = 50 μm. Representative images, n = 5 for each group. e–g Downregulated ATF6 expression in Pcdh20 CKO mice with DSS-induced colitis compared with WT mice by using immunofluorescence (e), semi-quantification of immunofluorescence with Image J software (f), and immunoblot (g). Magnification × 100, scale bars = 100 μm. Representative images (e). Each group n = 3. h Co-immunoprecipitation of PCDH20 (with flag-tag) and ATF6 in PCDH20-overexpressed Caco-2 cell line. i,j Paracellular permeability in polarized Caco-2 monolayer transfected with ATF6 overexpression plasmid or ATF6 shRNA. Transepithelial electrical resistance (TEER) (i), paracellular permeability assessment with FD4 (j). The data in f, i, j were presented as mean ± SEM. * p < 0.05 vs. Control, **p < 0.01 vs. Control