Figure 5.

Sirtuin 1 (SIRT1) mediated the acetylation of p53 and promoted autophagy in podocytes. (A) p53 acetylation was determined in Flag IP, and total p53 was determined on total extracts as input in podocytes cultured in high glucose for 48 h after transfection with si-SIRT1 or anti-miR-150-5p and expressing Flag-p53. (B) Acetylation and SIRT1 were determined in p53 IP, and total p53 was determined on total extracts as input in podocytes cultured in high glucose for 48 h after transfection with si-SIRT1 or anti-miR-150-5p. (C) The levels of p-AMPK, LC-3, and p62 in podocytes were determined using Western blot after transfection with si-SIRT1 and culture in high glucose for 48 h. (D,E) Typical images of immunofluorescence staining of mRFP-GFP-LC3 in podocytes cells after transfection with si-SIRT1 and culture in high glucose for 48 h. Typical profiles of autophagosomes (RFP + GFP + dots) and autolysosomes (RFP + GFP-dots) per cell section tested by confocal microscopy are shown and quantified. Data are expressed as mean ± SEM (n = 3; *p < 0.05, **p < 0.01, ***p < 0.001).