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Fig. 1 | Cancer & Metabolism

Fig. 1

From: Dynamic regulation of mitochondrial pyruvate metabolism is necessary for orthotopic pancreatic tumor growth

Fig. 1

Effect of PDHA1 engineering on PDC components and PDHA1 phosphorylation during hypoxia. A Schematic showing the flux of glucose-derived pyruvate in normoxia (top) and hypoxia (bottom). Note that in hypoxia, HIF1-induced expression of PDHK1/PDHK3 leads to hyperphosphorylation of PDHA1 and attenuated flux through PDH to produce acetyl-CoA for citrate production at citrate synthase. B Western blot of lysates from parental MiaPaca2 and engineered cells with the indicated alleles after growth in complete media after 16 h at either 21% or 1% oxygen showing consistent levels of E2 and E3, near complete loss of PDHB, and no change in VDAC and similar levels of phosphorylation of regulatory serine residues in parent and engineered wild type line. Note VDAC as a mitochondrial control, and beta tubulin for loading control. One of N=3 replicates is shown. C Western blot of parental MiaPaca2 and engineered cells grown in complete media for 16 h at either 21% or 1% oxygen as indicated. NB there appears to be sequential phosphorylation from 293 to 300 to 232 as mutation in upstream sites affects downstream efficiency of modification. One of N = 3 replicates is shown. AAA, PDHA1 with alanine substitutions at serines 232, 293, and 300. AAS, PDHA1 with an alanine substitution at serine 300. ASA, PDHA1 with an alanine substitution at serine 293. SAA, PDHA1 with an alanine substitution at serine 232

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