Fig. 2

Hypoxic response of compound mutant alleles of PDHA1. Western blot analysis of lysates from indicated engineered cell lines after 16 h in 21% or 1% oxygen. PDHA1 serine 232 phosphorylation is significantly reduced by single mutations of either 293 or 300, with almost a complete block when both are mutated. N.B. Mutation of all three sites completely abrogates hypoxic PDHA1 phosphorylation as detected by immunoreactivity. One of N = 3 replicates is shown. ASS, PDHA1 with alanine substitutions at serines 293 and 300. SAA, PDHA1 with alanine substitutions at serines 293 and 300