Figure 3
From: STAT3 Undergoes Acetylation-dependent Mitochondrial Translocation to Regulate Pyruvate Metabolism
STAT3 promotes the conversion of pyruvate to acetyl-CoA by PDC.
(a) Wild-type and STAT3−/− MEFs were treated with insulin for 30 min, and this was followed by immunoprecipitation of PDC E1. PDC E1 precipitates were subjected to western blotting for STAT3 co-immunoprecipitation. PDC E2 and PDC E3 were detected in the cell lysates. (b) Myc-STAT3 full length (FL) and Myc-STAT3 domains, as indicated, were cotransfected with PDC E1 in 293T cells, which were then subjected to by anti-Myc immunoprecipitation. STAT3 FL and STAT3 685-770 precipitated STAT3. (c) PDC E1 activity was measured in WT and STAT3−/− MEFs treated with insulin or left untreated. (d) 293T cells were transfected with an EV or Myc-tagged STAT3 fused with Cox4 MLS (Cox4-STAT3). Cytoplasmic (cyt) and mitochondrial lysates (mit) were analysed for Cox4-STAT3 acetylation and Cox4-STAT3 (anti-Myc) via western blotting. (e) Mitochondrial PDC E1 activity was measured in STAT3−/− MEFs transfected with an EV, Myc-STAT3, or Myc-Cox4-STAT3. (f) Mitochondrial PDC E1 activity was measured in STAT3−/− MEFs transfected with STAT3 WT, STAT3-3KR, STAT3-Y705F, or STAT3-S727A mutant. (g) Flag-STAT3 WT and Flag-STAT3 mutants, as indicated, were transiently transfected along with Myc-tagged PDC E1 in 293T cells. Anti-Myc immunoprecipitates were analysed for Flag-STAT3. (h) Acetyl-CoA content was measured in WT or STAT3−/− MEFs treated with insulin.
