Autophagy inhibition by Beclin1 siRNA also decreases the expression of podocin and impairs the filtration barrier of podocyte monolayer.

<p>(A): Western blot analysis shows that beclin-1 silencing which was achieved by using 20 pmol/L siRNA significantly inhibited podocin protein expression. Mouse podocytes were transfected with beclin-1 siRNA and then incubated for 24 hours. (B and C): Quantitative determination of beclin-1 protein (B) and podocin protein (C) abundance after normalization with α-tubulin. Data are presented as mean ± SEM of three independent experiments. *<i>P</i><0.05 vs. normal control; (D) Graphic presentation of the albumin influx across podocyte monolayer. Podocyts monolayer on collagen-coated Transwell filters was transfected with control siRNA or beclin-1 siRNA and then incubated for 24 hours, and albumin permeability across podocyte monolayer was determined. Values are means ± SEM; n = 3. *<i>P</i><0.05 vs. control; (E–G): Representative photographs of podocin visualized by indirect immunofluorescence staining in the control, the control siRNA transfected cells and the beclin-1 siRNA transfected cells. (E): control; (F): podocytes transfected with control siRNA for 24 hours; (G): podocytes transfected with beclin-1 siRNA (20 pmol/L) for 24 hours.