Figure 4.

Pan-PI3Ki augments chemotherapy and steroid-induced B-ALL cell death in vitro. (A and B) Proliferation of Nalm-6 and RCH-ACV cells treated with vehicle or copanlisib after three hour exposure to Ara-C or PBS. Data are mean ± s.e.m.; ANOVA with Tukey test at 72 h, n = 3 biologically independent experiments; p < 0.05, p < 0.01, p < 0.001, p < 0.0001. (C) Apoptosis in Nalm-6 cells as detected by Annexin V+Annexin V/PI or Annexin V/7-AAD. Cells were treated with vehicle or copanlisib alone or vehicle or copanlisib after three hour exposure to Ara-C. Data are mean ± s.e.m.; ANOVA with Tukey test, n 3 biologically independent experiments; p < 0.05, p < 0.01, p < 0.001, p < 0.0001. (D) β-catenin reporter GFP positivity detected over time by flow cytometry in Nalm-6 cells treated with vehicle or copanlisib after three hour exposure to Ara-C. Data are mean ± s.e.m.; unpaired two-sided Student’s t-test, n = 3 biologically independent experiments; p < 0.05, p < 0.01. (E–G) Proliferation of Nalm-6, RCH-ACV, or Sup-B15 cells treated with vehicle, GS-649443, or copanlisib alone or in combination with dexamethasone. Data are mean ± s.e.m.; ANOVA with Tukey test at 72 h, n = 3 biologically independent experiments; p <0.05, **p < 0.01, p < 0.001, p < 0.0001. (H) Apoptosis in Sup-B15 cells as detected by Annexin V+Annexin V/PI or Annexin V/7-AAD. Cells were treated with vehicle, GS-649443, or copanlisib alone or in combination with dexamethasone. Data are mean ± s.e.m.; ANOVA with Tukey test, n = 3 biologically independent experiments; p < 0.05, p < 0.01, p < 0.0001. (I) Proliferation of primary patient B-ALL cells treated with vehicle, GS-649443, or copanlisib. Data are mean ± s.e.m. (J) Apoptosis in primary patient B-ALL cells as detected by Annexin V+Annexin V/PI. Cells were treated vehicle, GS-649443, or copanlisib. Data are mean ± s.e.m.