Figure 4

GATA4 or GATA6 deletion in the jejunal epithelium perturbs enterocyte gene expression. (A) Hematoxylin and eosin (H&E) staining of jejunal tissue from controls and GATA4 or GATA6 mutants showed no difference in intestinal architecture. (B) Average jejunal villus length was determined using NDP Scan software (Hamamatsu) to measure the length of all villi per section. No difference was observed between controls and GATA4 or GATA6 mutants (G4 CTL, Gata4^loxP/+, n = 4 embryos, 3 sections/embryo, 166 villi, 129.2 ± 10.8 μm; G4 cKO, Gata4^loxP/-Villin-Cre, n = 3 embryos, 3 sections/ embryo, 132 villi, 144.1 ± 1.8 μm; G6 CTL, Gata6^loxP/+, n = 4 embryos, 3 sections/ embryo, 189 villi, 132.7 ± 6.6 μm; G6 cKO, Gata6^loxP/-Villin-Cre, n = 4 embryos, 3 sections/embryo, 173 villi, 127.7 ± 9.4 μm. Differences are not statistically significant) (C) Alkaline phosphatase (AP) staining was equivalent between controls and mutants. (D) qRT-PCR for jejunal-enriched enterocyte transcripts (Apoa4, Apoc2, Apoc3, Fabp1, Lct, Slc2a2, Slc2a5, Slc5a11) showed that 5/8 were decreased in GATA4 mutants and that 2/8 were decreased in GATA6 mutants. None of the transcripts expressed at similar levels in jejunum and ileum (Fabp2, Abcg5, Abcg8) were changed in either mutant compared with controls. (E) qRT-PCR for ileal-enriched enterocyte transcripts (Slc10a2, Fabp6, Fgf15, Cldn8) showed that expression of each was induced in jejunum in the absence of GATA4. Only Cldn8 was induced in jejunum in the absence of GATA6. For qRT-PCR in D and E, epithelial cells from three control (Gata4^loxP/+) and three mutant intestines (Gata4^loxP/-Villin-Cre or Gata6^loxP/-Villin-Cre) were assayed at least three times. Gapdh was used for normalization. All P-values were determined by two-sample Student’s t test: *P ≤ 0.05, **P ≤ 0.01. All error bars show SEM. Scale bars in all micrographs are 50 μm. All experiments performed using E18.5 embryos.