Suppression of autophagy in diabetic glomerular podocytes.

<p>(A) Western blot analysis shows the level of autophagy related proteins in diabetic glomerular. The glomerular lysates were blotted with specific antibodies against Beclin-1, Atg12-5 and LC3, respectively. Samples from two individual animals were used at each timepoint. (B) Quantitative determination of Beclin-1, Atg12-5 and LC3 protein abundance after normalization with α-tubulin. *<i>P</i><0.05 (n = 3). (C–D) Immunofluorescence staining shows the changes of autophagosomes in the diabetic glomerular (400× magnification). Kidney section were immunostained with anti-LC3 antibody (green) to identify autophagosomes, followed by staining with anti-Laminin antibody (red) to sever as a marker for the basement membrane.(C, E and G): control group; (D, F and H): the group of 28 day diabetic mouse. (I): Quantification of LC3 immunofluorescence staining in nondiabetic glomeruli and diabetic glomeruli. 30 glomeruli were evaluated for each experimental animal (n = 5) through the middle part of the kidney. Glomeruli that did not have a glomerular tuft or that were sectioned close to the edge were disregarded. Data are presented as mean ± SEM. *<i>P</i><0.05 vs. normal control.