Fig. 5: Deacetylase SIRT5 deacetylates MRPL12 at K163.
From: MRPL12 K163 acetylation inhibits ccRCC via driving mitochondrial metabolic reprogramming
A Immunoblot (IB) analysis of acetylation, stably expressed Flag-MRPL12 in HEK293T cells treated with TSA (5 μM for 16 h) and NAM (10 mM for 6 h). B IB analysis of acetylation, stably expressed MRPL12 in OS-RC-2 and 786-O cells treated with TSA (5 μM for 16 h) and NAM (10 mM for 6 h). C Analysis of MRPL12 K163 acetylation levels in OS-RC-2 cells transfected with siRNA targeting SIRT deacetylases. D Endogenous interaction between MRPL12 and SIRT5 in OS-RC-2 and 786-O cells, determined by immunoprecipitation (IP) and IB analysis. E HEK293T cells transfected with Flag-MRPL12 and HA-SIRT5 as indicated. Interactions between MRPL12 and SIRT5 were determined by co-IP and IB analysis. F AlphaFold 3 simulation showing the predicted interaction between MRPL12 and SIRT5. G Duolink proximity ligation assay demonstrating the interaction between MRPL12 and SIRT5 in OS-RC-2 cells. Scale bar: 10 μm, n = 3 per group. H Endogenous MRPL12 K163 acetylation increases upon SIRT5 silencing. I Endogenous MRPL12 K163 acetylation decreases with SIRT5 overexpression. J Effect of SIRT5 knockdown and overexpression on cell migration in OS-RC-2 cells. Scale bar: 100 μm. K SIRT5 expression levels in clear cell renal cell carcinoma (ccRCC) tissues compared with normal tissue (NTL) tissues, as shown in the UALCAN database, indicating reduced expression in ccRCC. Significance levels: ns, P > 0.05; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.
