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

Fig 1. KSHV infection promotes cell proliferation and DNA replication in infected cells grown under hypoxic conditions.

Fig 1

(A) Confirmation of induction of hypoxia by western blot for LANA and PDK1. (B) Cell cycle analysis of BJAB and BJAB-KSHV cells grown under normoxic or 1% O2 induced hypoxic conditions. Cells were grown for under normoxic or hypoxic conditions and stained by propidium iodide after fixation and RNase treatment. Representative image for cell cycle analysis at 24 hours are shown. (C) Barr diagram showing relative percentage of cells in various cell cycle phases in BJAB and BJAB-KSHV cells grown under normoxic or 1% O2 induced hypoxic conditions. (D) Schematic showing cyclin/cyclin dependent kinase pair involved in various stage of cell cycle. (E, F and G) Real-time PCR expression analysis of Cyclin D1 (CCND1), Cyclin E (CCNE) and cyclin dependent kinase 2 (CDK2) in BJAB and BJAB-KSHV cells grown under normoxic or 1% O2 induced hypoxic conditions. Cells were grown for 24 or 36 hours in normoxic or 1% O2 induced hypoxic conditions followed by RNA isolation and cDNA synthesis. Relative fold change was calculated by delta CT method taking GAPDH as endogenous control. (H) Western blot analysis for the CyclinD1, Cyclin E and CDK2 in BJAB and BJAB-KSHV cells grown under normoxic or 1% O2 induced hypoxic conditions. Cells were grown under normoxic or 1% O2 induced hypoxic conditions for 36 hours and the whole cell lysate was used for probing protein levels as indicated. GAPDH served as endogenous control. (I) Western blot analysis for the CyclinD1, Cyclin E and CDK2 in HEK293T and HEK293T-BAC16-KSHV cells grown under normoxic or 1% O2 induced hypoxic conditions. Cells were grown under normoxic or 1% O2 induced hypoxic conditions for 24 hours and the whole cell lysate was used for probing protein levels as indicated. GAPDH served as endogenous control.