Fig. 7
Effects of Ap4 deletion on intestinal organoids. a qPCR analysis of the indicated mRNA from organoids isolated from three female mice per genotype after passaging and 7 days after Cre activation by 4-OHT. b Left panel: representative pictures of small intestinal organoids 7 days after passaging and Cre activation by 4-OHT. Scale bars represent 20 µm. Right panel: quantification of protrusions (crypt-like structures) per organoid derived from three female mice per genotype. A total of 81 organoids were evaluated per genotype. c Venn diagram displaying differentially regulated mRNAs (fold change ≥ 1.5, p < 0.05) in Ap4fl/fl and Ap4∆IEC organoids as determined by edgeR and DESeq2. d Volcano plot and heatmap depicting expression changes of differentially expressed mRNAs between Vil-Cre-ERT2 and Vil-Cre-ERT2/Ap4fl/fl organoids, 7 days after Cre activation by 4-OHT, detected by RNA-Seq. Left panel: volcano plot depicting expression changes of differentially expressed mRNAs (fold change ≥ 1.5) from Ap4fl/fl and Ap4∆IEC organoids. Downregulated mRNAs are depicted in blue, upregulated mRNAs are depicted in red. RNAs with fold changes < 1.5 and/or statistically nonsignificant changes in expression are indicated in black. Dashed vertical lines indicate 1.5-fold change cutoff. Dashed horizontal line indicates the cutoff for adjusted p-values < 0.05 as determined with DESeq2. Right panel: Heatmap depicting expression changes of differentially expressed mRNAs (fold change ≥ 1.5 p < 0.05 as determined by edgeR and DESeq2) from Ap4fl/fl and Ap4∆IEC organoids. Colors indicate relative expression values from minimum (blue) to maximum (red) for each RNA sample per differentially regulated mRNA. a, b Results represent the mean ± SD. Results were subjected to an unpaired, two-tailed Student’s t-test with p-values * < 0.05, ** < 0.01, *** < 0.001, n.s.: not significant
