GATA4 or GATA6 deletion in the jejunal epithelium alters goblet cell number and enteroendocrine cell gene expression. (A) MUCIN 2 (MUC2) IHC indicated a subtle increase in goblet cells in Gata4 and Gata6 cKO jejunum compared with controls. (B) Jejunum from Gata4 cKO and Gata6 cKO embryos contained more goblet cells than control jejunum (G4 CTL, Gata4loxP/+, 68 ± 4 MUC2+ cells/section; G4 cKO, Gata4loxP/-Villin-Cre, 100 ± 10 MUC2+ cells/section; G6 CTL, Gata6loxP/+, 75 ± 8 MUC2+ cells/section; G6 cKO, Gata6loxP/-Villin-Cre, 100 ± 1 MUC2+ cells/section; n = 3 embryos/genotype, 3 sections/embryo) (C) Chromogranin A (CHGA) IHC appeared comparable among jejunum from Gata4 cKO, Gata6 cKO, and control embryos. (D) qRT-PCR for enteroendocrine cell markers showed that pan-enteroendocrine cell marker expression (Ngn3, ChgA) was unchanged in the absence of GATA4 or GATA6. Expression of proximal enteroendocrine cell markers was decreased in GATA4 mutant jejunal epithelium (Gip, Cck). Expression of Pyy, a distally-enriched enteroendocrine cell marker, was increased in GATA4 mutants and decreased in GATA6 mutants. Epithelial cells from three control (Gata4loxP/+) and three mutant intestines (Gata4loxP/-Villin-Cre or Gata6loxP/-Villin-Cre) were assayed at least three times. Gapdh was used for normalization. (E) SOX9 IHC was unchanged between knockouts and controls. (F) Loss of GATA4 or GATA6 did not alter SOX9+ cell number in the jejunum (G4 CTL, Gata4loxP/+, 367 ± 44 SOX9 + cells/section; G4 cKO, Gata4loxP/-Villin-Cre, 367 ± 5 SOX9 + cells/section; G6 CTL, Gata6loxP/+, 299 ± 25 SOX9 + cells/section; G6 cKO, Gata6loxP/-Villin-Cre, 343 ± 21 SOX9 + cells/section; n = 3 embryos/genotype, 3 sections/embryo; Differences were not statistically significant). P-values were determined by two-sample Student’s t test: *P ≤ 0.05, **P ≤ 0.01, #P = 0.0744. Error bars show SEM. Scale bars are 50 μm. All experiments performed using E18.5 embryos.