Fig. 3: Analysis of HIF1α interactions and TLS across DNA lesions under hypoxia.
From: Hypoxia-dependent recruitment of error-prone DNA polymerases to genome replication
A HEK293FT cells were transfected with the indicated siRNA (50 nM) and incubated for 48 h, followed by 24 h incubation under hypoxia (0.5% oxygen) after which chromatin-bound and soluble proteins A were analyzed by immunoblotting for the presence of the indicated proteins, as described above. B HEK293FT cells were transfected with HA-HIF1αP402A/P564A expressing plasmid or the empty vector for 24 h. Proteins were then extracted and immunoprecipitated (20μg) with an anti-HA antibody, followed by SDS-PAGE and blotting with the indicated antibodies. C–E Cells were subjected to hypoxia or normoxia for 24 h after which they were assayed for TLS activity using the TLS gap-lesion plasmid assay. C The effect of hypoxia on TLS across a cisPt-GG or BP-G lesion in HEK293FT cells. D The effect of hypoxia on TLS across a cisPt-GG lesion in MCF7 (breast cancer) and A549 (lung cancer) cells. E TLS mutagenicity under hypoxia was assessed by sequencing the region filled-in by TLS in gap-lesion plasmids with a cisPt-GG or BP-G adduct. The detailed sequence data is presented in Supplementary Table S1.
