Fig. 6

PAF1 regulates LDHA transcription through HIF1α binding. A Consensus Human HIF1α binding motif generated on https://jaspar.genereg.net/. B LDHA promoter sequence showing three HIF1α binding sites. C ChIP assay was performed using PAF1 and control IgG antibodies in MiaPaCa2 cells using primers that amplify LDHA promoter, which contains HIF1α binding motif. Immunoprecipitated and purified DNA was amplified using PCR followed by running in agarose gel. Binding site 2 and 3 did not amplify. D ChIP Re-ChIP assay was performed using PAF1, HIF1α and control IgG antibodies in MiaPaCa2 cells using primers that amplify LDHA promoter. Immunoprecipitated and purified DNA was amplified using PCR followed by running in agarose gel. Binding sites 2 and 3 did not amplify E ChIP assay was performed using PAF1 and control IgG antibodies in MiaPaCa2 shPAF1 cells using primers that amplify LDHA promoter for B.S.1. Immunoprecipitated and purified DNA was amplified using PCR followed by running in agarose gel. Binding site 1 did not amplify. F Representative sequence generated from PCR amplification and purification, with HIF1α binding motif highlighted. G Schematic shows the overall summary of the study