Fig 5. Bcl-xL is not required for survival during latent infection in other cell types.
(A) BCBL-1, cells were transduced with sgRNAs selected for using puromycin resistance for 4 days, then seeded at 1 x 105 cells/mL. Cell viability was measured using a trypan blue assay at 24, 48, 72, and 96 h post seeding. (B) BCBL-1 cells were seeded at an initial concentration of 1 x 105 cells/mL and growth media was supplemented with or without A-133 at 1 nM or 10 nM. Cell viability was measured using a trypan blue assay at 24, 48, 72, and 96 hpi. (C) BC-1 or (D) JSC-1 were treated with A-133 as in (B). (E) BJABs were transduced with sgRNAs and treated as in (A). (F) 293T cells or (G) HFF cells were transduced and selected for sgRNAs to Bcl-xL. Transduced cells were mock or KSHV infected and cell viability was measured using a trypan blue assay 48 hpi (F) or measured by counting live (Syto59) and dead (YoYo-1) cells using Cell-Cyte (G). (H) Western blot analysis with antibodies to Bcl-xL, Mcl-1, and Bcl-2 in cell lines tested above. Data are presented as mean ± s.d. (2-way ANOVA, n = 3) *P<0.05.