Fig. 6

EMP3 KOs have impaired proliferative response and increased sensitivity to kinase inhibition. A, B Proliferation of DK-MG (A) and U-118 (B) control and EMP3 KO cells cultured in normal serum-containing medium over the course of 4 days. Dots represent mean fold-changes in CellTiter-Glo luminescence readings relative to t = 0, while error bars show 95% confidence intervals (unpaired t-test; ***P < 0.001, ****P < 0.0001). C, D Proliferative response of serum-starved DK-MG (C) and U-118 (D) control and EMP3 KO cells to daily EGF treatment for 72 h. Bar plots represented mean fold-changes in CellTiter-Glo luminescence readings of EGF-treated cells versus untreated controls. Error bars and dots represent S.D. and individual measurements, respectively (unpaired t-test; **P < 0.01, ****P < 0.0001). E, F Caspase 3/7 levels in DK-MG (E) and U-118 (F) control and EMP3 KO cells treated with 1 μM staurosporine (STS) for 4 h. Bar plots represent mean fold-changes in Caspase 3/7 luminescence readings of STS-treated cells versus untreated controls. Error bars and dots represent S.D. and individual measurements, respectively (Welch’s t-test; ***P < 0.001, ****P < 0.0001). G Western blot showing full-length (green arrow) and cleaved PARP (magenta arrow) in U-118 control and EMP3 KO cells treated with 100 nM or 1 μM STS for 4 h. H Concentration–response curve of U-118 control, EMP3 KO, and EMP3 KO + EMP3 cells treated with the EGFR inhibitor AZD9291 for 24 h. Points represent mean percentage changes in CellTiter-Glo luminescence levels of treated cells relative to untreated cells, while error bars represent S.D. (n = 3). I Caspase 3/7 levels in U-118 control, EMP3 KO, and EMP3 KO + EMP3 cells treated with 2.5 μM AZD9291 for 24 h. Bars represent mean fold-changes in Caspase 3/7 luminescence readings of treated cells versus untreated controls. Error bars and dots represent S.D. and individual measurements, respectively (Welch’s ANOVA with Dunnett’s T3 multiple comparisons test; *P < 0.05)