Fig. 3.

mTOR enhances PGAM1 expression through induction of HIF1α. Immunoblots of wt and Tsc2^−/− a or Pten^−/− b MEFs transduced with shHIF1α or scramble shRNA lentiviruses. c Schematic representation of the promoter regions of mouse PGAM1 gene. Site 1 and site 2 indicate the potential binding sequences of HIF1α on PGAM1 promoter. Predicted binding region (PBR) for real-time PCR covers both binding sites. The transcription start site is indicated by an arrow above the gene. d Tsc2^−/− MEFs was treated with or without 10 nM rapamycin (Rapa) for 24 h. HIF1α antibody-immunoprecipitated DNA from wt and Tsc2^−/− MEFs was PCR amplified for PBR and NBR regions. The data are plotted as the ratio of immunoprecipitated DNA subtracting nonspecific binding to IgG vs. total input DNA. Representative data from two independent experiments are shown. Data represent mean ± SEM of replicate real-time PCR. **P < 0.001