Figure 1: RNAi-mediated hepatic silencing of Itgb1.
(a) Dose-dependent Itgb1 mRNA knockdown 3 days after siRNA injection(n=4–5, mean±s.e.m., P-levels—comparison to the si-Control-treated group by the Turkey post hoc test, (b) time-course of Itgb1 knockdown on the mRNA and protein level after a single dose of siRNA (0.5 mg kg−1, n=3). P<0.001 for mRNA levels in si-Itgb1-treated animals (time points, 3–24 days compared with si-control-treated animals by the Turkey post hoc test. P-levels on the graph for comparison of Itgb1 protein level with si-control-treated animals by the Turkey post hoc test. (c) Representative western blot of the time-course. (d) Analysis by flow cytometry of the Itgb1 expression on hepatocytes isolated from animals treated twice with Itgb1 siRNA. (±1° Ab—isotype control/primary antibodies added). Number indicates geometry mean of fluorescent intensity of si-Itgb1-treated cells normalized to one of the control group and represented as mean±s.e.m. For extensive analysis of expression of other integrin subunits, their quantification Supplementary Fig. 2 and for mRNA knockdown quantification see Supplementary Fig. 2d. (e) Analysis of Itgb1 expression by immunofluorescent staining, expression on hepatocytes (red arrows, lateral surface and white arrow, basal surface). Scale bar, 20 μm.
