Fig. 7.

SIRT3 deacetylates PRDX3 at K253 in Caco-2 cells.

(A) PRDX3 acetylation in Caco-2 cells transfected with potential acetylation site-specific mutants of PRDX3, n = 3. (B) Partial amino acid fragment of PRDX3 in several species. (C, D) PRDX3 acetylation, n = 3. (E–L) Caco-2 cells were transfected with no-load plasmid, PRDX3 expression plasmid, PRDX3 K253R or K253Q plasmid and then subjected to H/R injury; these cells groups are denoted pcDNA 3.1, K253WT, K253R and K253Q, respectively. (E) Representative Western blot of PRDX3 monomer and dimer in Caco-2 cells H/R injury. (F) Mitochondrial H₂O₂ level, n = 8. (G) MitoSOX Red and flow cytometry analysis of cells stained with MitoSOX dye. Scale bar = 25 μm, n = 6. (H) The mitochondrial membrane potential was measured by JC-1, and the ratio of red to green fluorescence ratio in isolated mitochondria stained with JC-1 probes was calculated. Scale bar = 50 μm, n = 6. (I) The mitochondrial mass was measured by immunoblotting of TOMM20, n = 3. (J) The mitochondrial morphology was determined by TEM. Scale bar = 10 μm. (K) The expression of cleaved caspase-3 protein was normalized to β-actin expression in total cell lysates from Caco-2 cells, n = 3. (L) Caspase-3 activity in Caco-2 cells, n = 8. (M) TUNEL and DAPI staining. The apoptotic index is represented as the ratio of TUNEL-positive cells to DAPI staining. Scale bar = 50 μm, n = 6. *p < 0.05, **p < 0.01. . (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)