Fig 2: qPCR characterization of primary cIECs and cIEC-H2 passage 4 pre- and post-mEGF differentiation. Data are reported as fold change calculated using the ??CT method. RNA samples with a 260/280 ratio below 2.0 were excluded from the analysis. Outliers were defined by the ROUT Method with the False Discovery Rate (Q) set >1% and excluded from the analysis. Data were normally distributed (Shapiro–Wilk test P > 0.05) and analyzed using one-way ANOVA with Tukey's multiple comparisons. CDH1: undifferentiated vs. differentiated P < 0.0001, 95% CI -1.848 to -1.074, differentiated vs. primary P < 0.0001, 95% CI 0.9348–1.933—EPCAM undifferentiated vs. differentiated P < 0.0001, 95% CI -8.590 to -4.591, undifferentiated vs. primary P = 0.0348, 95% CI -5.204 to -0.2155, differentiated vs. primary P = 0.0066, 95% CI 1,299–6,462—VIM: undifferentiated vs. differentiated P < 0.0001, 95% CI -12.90 to -10.54, differentiated vs. primary P < 0.0001, 95% CI 10.68–14.76—KRT18 undifferentiated vs. differentiated P < 0.0001, 95% CI 0.3084–0.6180, undifferentiated vs. primary P = 0.0034, 95% CI -0.5174 to -0.1313, differentiated vs. primary P = 0.0066, 95% CI -0.9874 to -0.5877—KRT20: undifferentiated vs. differentiated P = 0.0009, 95% CI -95.54 to -33.69, differentiated vs. primary P = 0.0048, 95% CI 23.35 to 103.2– ALPi: undifferentiated vs. primary P = 0.0026, 95% CI 0.4351–1.381, differentiated vs. primary P = 0.0163, 95% CI 0.1393–1.036—ZO1: undifferentiated vs. primary P = 0.0165, 95% CI -1.467 to -0.1825, differentiated vs. primary P = 0.0073, 95% CI -1,612 to -0.3270—OCLN: undifferentiated vs. differentiated P < 0.0001, 95% CI -1.066 to -0.6087, differentiated vs. primary P = 0.0008, 95% CI 0.3244 to 0.9145—CLDN1: undifferentiated vs. differentiated P < 0.0001, 95% CI -1.718 to -0.8292, undifferentiated vs. primary P = 0.0196, 95% CI 0.1251–1.234, differentiated vs. primary P < 0.0001, 95% CI 1.379–2.527. Data are represented as mean with SEM (n = 5). Abbreviations: ALPi = intestinal alkaline phosphatase; CDH1 = E-cadherin; CLDN1 = claudin 1; EPCAM = epithelial cellular adhesion molecule; KRT18 = cytokeratin 18; KRT20 = cytokeratin 20; KRT8 = cytokeratin 8; MAGM = maltase-glucoamylase; OCLN = occludin; SUC-2 = intestinal sucrase-isomaltase; VIL1 = villin; VIM =vimentin; ZO1= zonula occludens 1.