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. 2019 Sep 3;15(9):e1008025. doi: 10.1371/journal.ppat.1008025

Fig 7. HIF1α is necessary for LANA-mediated protection of replication associated proteins from hypoxia-dependent degradation.

Fig 7

(A) BJAB and BJAB-KSHV cells were transfected with plasmid vector encoding ShControl or ShHIF1α. The HIF1α knock-down was confirmed by real-time PCR. Briefly, 48 hours post transfection, total RNA was isolated and cDNA was synthesized. The cDNA was used for real-time PCR. (B) HIF1α knock down was confirmed by calculating fold change expression of a known HIF1α target P4HA1 by growing these cells under hypoxic conditions. The experiments were performed at least in triplicate. The error bar represents standard error from the mean. Asterisk represents statistically significant difference. (C) Representative image for Western blot analysis of replication-associated proteins in BJAB-ShControl, BJAB-ShHIF1α, BJAB-KSHV-ShControl and BJAB-KSHV-ShHIF1α cells grown under normoxic or 1% O2 induced hypoxic conditions. The ShControl or ShHIF1α cells were grown under normoxic or 1% O2 induced hypoxic conditions for 24 hours. Equal amounts of protein were used to probe with indicated antibodies. (D) Representative image for Western blot analysis of replication associate proteins in BC3-ShControl, BC3-ShHIF1α cells grown under normoxic or 1% O2 induced hypoxic conditions. The ShControl or ShHIF1α expressing cells were grown under normoxic or 1% O2 induced hypoxic conditions for 24 hours. Equal amounts of protein were used to probe with indicated antibodies. GAPDH served as loading control for panels C and D.