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. 2015 Aug 18;17(7):564–573. doi: 10.1016/j.neo.2015.07.003

Figure 4.

Figure 4

Knockdown of ATF5 has no effect on MCL1 expression and apoptosis sensitivity. Verification of the ATF5 knockdown in LN229 and U87 cells as analyzed by qPCR (A). Two single-cell clones per cell line are shown. Data represent means of n = 3 cultures + SEM; *P < .05, significant difference to respective control cell line. MCL1 expression of ATF5 knockdown cells in comparison to LN229 control cells (Ø) as analyzed by Western blotting. GAPDH served as loading control. mRNA levels of MCL1 were assayed by qPCR in control cells with the corresponding knockdown clones of ATF5 and MCL1 in LN229 (B, left panel) and in U87 (C, left panel) +/− treatment with 5 μM sorafenib for 24 hours. Data represent means of n = 3 cultures + SEM; *P < .05, significant difference to respective control cell line. Experiments were repeated three times with similar results. Effector caspase activity after treatment for 4 hours with 5 μM sorafenib, 5 μM ABT-737, or the combination of both (C, D, right panels). Experiments were repeated three times with similar results. All graphs represent means of n = 4 cultures + SEM; *P < .05, significant difference to untreated controls; #P < .05, significant difference to respective control cell line. n.s.: not significant.