Fig 1: Western blot analysis of ACO2, CES2 and ANXA3A. Western blot images of ACO2, CES2 and ANXA3. ß-actin was used as loading control. B. Relative protein abundances determined by western blot (Mean±standard deviation). Abbreviations: N, normal colonic mucosa; A, adenoma; C, colorectal cancer.
Fig 2: CDK2-mediated phosphorylation destabilizes PXR protein and suppresses PXR-targeted genes expression. (A,B) HepG2 cells were treated with okadaic acid (0.1 µM) for 24 h, and the gene expression was analyzed at protein (A) or mRNA levels (B). Cell lysates were prepared and subjected to Western blot to determine the expression of PXR, CES1, CES2, CYP3A4 and P-gp, respectively. mRNA levels were detected by qRT-PCR. (C,D) HepG2 cells were transfected with si-CDK2 or si-CDK5 and siRNA-control for 48 h. Cell lysates were prepared and subjected to Western blot to determine the expression of PXR, CES1, CES2, CYP3A4 and P-gp, respectively. (E) The effect of CDK2 overexpression on the suppression of CES1, CES2, PXR, CYP3A4 and P-gp. Experiments described in this figure were repeated independently at least three times, and data are expressed as mean ± SEM (n = 3). * p < 0.05 indicates significant difference compared with control group.
Fig 3: Immunohistochemistry images of S100A9, ANXA3, NAMPT, CES2 and CLCA1
Fig 4: Divergence of drug absorption in ileum. A, Expression heatmap of signature genes in enterocytes and functional enrichments among five species. B, Pearson correlation of scRNA-seq expression (log1p (average expression)) for drug metabolism and transporter genes. Species from top to bottom: macaque versus human, pig versus human, rat versus human and mouse versus human. C, Violin plots showing the expression of drug metabolism and drug transporter genes across human, macaque, pig, rat and mouse. ND, not detected. D, Immunofluorescence was performed to confirm the CES1 and CES2 expression in macaque, pig and human ileum. Scale bars, 100 µm. E, The expression of CYP3A family members in differentiated mouse, pig and human ileum organoids relative to ß-actin, and its activity was examined by P450-Glo CYP3A4 assay kit. F SLC22A1 expression relative to ß-actin and metformin uptake in mouse, pig and human ileum organoids. Data are displayed as the mean ± SD (n = 3 independent experiments). ns (no significance); *P < 0.05, **P < 0.01, ***P < 0.001 by Student’s t-test (E and F)
Supplier Page from Abcam for Anti-CES2 antibody [EPR14856]