Fig. 3.
EMA401 significantly inhibits the growth of GBM cells via AT2R. (A) TB77 cells were treated with 1, 10, and 30 μM PD123319 (PD) or EMA401 (EMA) in 1% serum and analyzed using the CCK8 kit 9 d posttreatment. (B) Primary GBM explants GBM31, TB26, TB43, TB48, GBM59, and GBM96 were treated with 1, 10, and 30 μM EMA401 in 1% serum and analyzed 9 d posttreatment with CCK8. Data shown are mean absorbance A450 values normalized to the untreated control (UC) ± SEM (C) dCas9-KRAB inducibility in 8MG-KRAB cells treated with 0.1, 0.5, 1, and 2 μg/mL doxycycline (Dox) for 48 h. Induction of dCas9-KRAB in 8MG-KRAB cells with stable integration of AGTR2 single-guide RNA (sgRNA) results in significant AGTR2 transcriptional silencing in the presence of 2 μg/mL doxycycline. Expression data were normalized to the mean cycle threshold value of the reference genes and presented as 2-ΔΔCt. (D) 8MG-KRAB cells with stable integration of scramble control (CTL) sgRNA or AGTR2 sgRNA were treated with 10 nM AngII, 30 μM PD123319, and 30 μM EMA401 in the presence of 2 μg/mL doxycycline in 1% serum. Cells were analyzed 9 d posttreatment by SRB staining. Data shown are mean absorbance A490 values normalized to the untreated control ± SEM. For all data presented in this figure, ANOVA with Tukey’s multiple comparisons test was performed to determine significant differences between groups. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. ns, nonsignificant.