Confirmation of intestinal KO of Heph in adult experimental mice. Genotypes of mice were determined by PCR amplification of the Cre and T-cell receptor sequences from genomic DNA isolated from tail tips followed by agarose gel electrophoresis. The amplicon at ∼1100 bp is the Cre transgene, and the band at ∼230 bp is the T-cell receptor (which was used as a positive control) (A). Expression of Heph mRNA in duodenal enterocytes was assessed by quantitative reverse transcription (qRT)-PCR and normalized to the expression of cyclophilin A (which did not vary significantly between samples) (B). n = 6 mice per group, with duplicate technical replicates. KO of intestinal Heph was also confirmed at the protein level by western blotting (C). The blot shown is representative of 6 independent experiments with similar results. Data are shown as box-and-whisker plots and were analyzed by 2-way ANOVA (B-C). A genotype main effect was noted in regards to Heph mRNA and protein expression (P < .001 for both). No sex main effects or 2-way interactions were noted. Ctrl, control.