CRISPR/CAS9-mediated loss of BCL-W does not effect the survival of select BL and DLBCL cell lines. (A) Cell viability following doxycycline induction of nontargeting (sgNT) or BCL-W–targeting (sgBCL-W) sgRNAs over 96 hours. Cell viability was measured by the proportion of cells that were Annexin V and DAPI double negative by flow cytometry. Error bars represent the SD for 3 independent experiments. (B) Sequencing results for cell lines 5 days and >1 month postinduction of BCL-W sgRNA. Bars show the proportions of reads which were wild type, in frame indels, or frameshift in the BCL-W gene. The number of reads for each cell line is indicated. (C) Western blotting for prosurvival BCL-2 family members and proapoptotic BIM in sgBCL-W and control (sgNT) Ramos-BL and HT cells at 5 days and >1 month post-sgRNA induction. Probing for HSP70 was used as a loading control. (D) BCL-2 family gene expression in sgBCL-W and sgNT control Ramos-BL and HT cells as determined by qRT-PCR analysis. Data are normalized to HMBS housekeeping gene expression levels and shown relative to HMBS levels. Error bars represent the SD of 2 independent experiments. Significance is indicated (Student t test, *P < .05).