Figure 4.
Figure 4. IKZF1, IKZF3, and ZFP91 are dispensable for the survival of PEL cell lines. (A-B) Growth curve analyses of Cas9-expressing BC-3 or BCBL-1 cell clones after transduction with lentiviral sgRNA vectors targeting IKZF1 (A) or IKZF3 (B). Two independent sgRNAs (sg1 and sg2) were designed per target; sgRNAs targeting AAVS1 and IRF4 were used as negative and positive controls, respectively (n = 3; error bars represent standard error of the mean [SEM]). Representative western blot analyses below the growth curves confirm the efficient inactivation of the targeted genes and demonstrate lack of regulation of IRF4. GAPDH served as loading control. (C) Growth curve analyses of naïve BC-3 or BCBL-1 cells after transduction with lentiviral shRNA vectors targeting IKZF1 or IKZF3. The nontargeting shRNA NT4 served as negative control (n = 3; error bars represent SEM). Representative western blot analyses below the growth curves assess the degree of shRNA-mediated target knockdown and IRF4 expression. GAPDH served as loading control. (D) Same as panels A and B, except that sgRNAs against ZFP91 were used (n = 3; error bars represent SEM). KO, knockout.

IKZF1, IKZF3, and ZFP91 are dispensable for the survival of PEL cell lines. (A-B) Growth curve analyses of Cas9-expressing BC-3 or BCBL-1 cell clones after transduction with lentiviral sgRNA vectors targeting IKZF1 (A) or IKZF3 (B). Two independent sgRNAs (sg1 and sg2) were designed per target; sgRNAs targeting AAVS1 and IRF4 were used as negative and positive controls, respectively (n = 3; error bars represent standard error of the mean [SEM]). Representative western blot analyses below the growth curves confirm the efficient inactivation of the targeted genes and demonstrate lack of regulation of IRF4. GAPDH served as loading control. (C) Growth curve analyses of naïve BC-3 or BCBL-1 cells after transduction with lentiviral shRNA vectors targeting IKZF1 or IKZF3. The nontargeting shRNA NT4 served as negative control (n = 3; error bars represent SEM). Representative western blot analyses below the growth curves assess the degree of shRNA-mediated target knockdown and IRF4 expression. GAPDH served as loading control. (D) Same as panels A and B, except that sgRNAs against ZFP91 were used (n = 3; error bars represent SEM). KO, knockout.

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