Figure 3. Genetic manipulations can alter PDH E1α phosphorylation in response to hypoxia.

(a) MIA PaCa-2 control (Hg), silenced HIF1α (shHIF1α) or silenced PDHK1 (shPDHK1) and PDP1 overexpressing cells were incubated for 16 h in normoxia or hypoxia (0.5% O₂) and phospho-serine E1α detected by Western blot showing that all three modifications can have an inhibitory effect on hypoxic Ser232 phosphorylation. (b) Increased pSer232-E1α detection after E1α immunoprecipitation from lysates of cells treated as in (a). (c) The same analysis as in (a) after treatment with 16 h 1 mM DMOG.