Effect of chronic hypoxia (1% O2), acute hypoxia (1% O2), and acute hypoxia plus TPA on KSHV intracellular lytic protein expression in BC-3 and BCBL-1 cells 48 to 72 hours after treatment.
Cells were seeded at 300 000 viable cells per milliliter. (A, B) Cells were maintained in either 21% O2 or 1% O2 for at least 3 passages before being used in experiments (chronic exposure). The plotted values for PF-8 (░) and gpK8.1 (▨) expression represent the median of at least 3 independent experiments ± the 25% and 75% quartiles. Exact Wilcoxon rank sum test (unpaired) for panel A: (BC-3, PF-8 21% O2 vs 1% O2) p2 =0 .0012, and (BC-3, gpK8.1 21% O2 vs 1% O2) p2 = 0.016. Exact Wilcoxon rank sum test (unpaired) for panel B: (BCBl-1, PF-8 21% O2 vs 1% O2) p2 = 0.004, and (BCBL-1, gpK8.1 21% O2 vs 1% O2)p2 = 0.028. (C, D) Cells grown in normoxia were incubated in 21% O2 or 1% O2 overnight. They were then exposed to TPA (6.4 nM) or to media control and returned to their respective conditions. The plotted values for PF-8 expression is the median percentage of positive cells for 6 independent experiments ± the 25% and 75% quartiles. Wilcoxon signed rank test (paired) for panel C: (BC-3, PF-8 21% O2 vs 1% O2)p2 = 0.031, and (BC-3/TPA, PF-8 21% O2 vs 1% O2) p2 = 0.031. Wilcoxon signed rank test (paired) for panel D: (BCBL-1, PF-8 21% O2 vs 1% O2) p2 = 0.031, and (BCBL-1/TPA, PF-8 21% O2 vs 1% O2) p2 = 0.031.
