1. Fig. 5 BRD0705 induces differentiation in AML cell lines and primary patient samples through GSK3α-selective inhibition.
BRD0705 induces differentiation in AML cell lines and primary patient samples through GSK3α-selective inhibition. (A and B) Time-response (A) and dose-response (B) Western immunoblots for β-catenin, phospho-GSK3α/β (Tyr279/216), total GSK3α/β, glycogen synthase (GYS), phospho-GYS (S641), and vinculin after treatment with the indicated inhibitors in U937 cells. (C) Dose-response Western immunoblot for β-catenin, phospho-GSK3α/β (Tyr279/216), total GSK3α/β, and vinculin after treatment with the indicated inhibitors in primary AML patient samples. (D) May-Grunwald-Giemsa staining after BRD0705 treatment for 6 days. Scale bar, 10 μm. (E) Fluorescence-activated cell sorting (FACS) analysis of the expression of CD11b, CD11c, and CD14 cell surface markers after BRD0705 treatment. *P < 0.05 calculated using a Welch’s t test in comparison with the control condition. Data are means ± SEM of three replicates. (F) FACS analysis of the expression of CD14 and CD117 cell surface markers in five primary AML samples treated with BRD0705. *P < 0.05 calculated using nonparametric Mann-Whitney test in comparison with control condition. Data are means ± SEM of five primary AML samples. (G) Western immunoblot for total GSK3α/β and vinculin in U937 after GSK3α CRISPR KO. (H) FACS analysis of the expression of CD11b cell surface marker in U937 GSK3α-WT, GSK3α-KO#1, and GSK3α-KO#2 treated with the indicated inhibitor. P < 0.05 calculated using a Welch’s t test in comparison with the DMSO condition for each clone (#) or the DMSO condition in the GSK3α-WT clone (*). Data are means ± SEM of three replicates.
Florence F. Wagner et al., Sci Transl Med 2018;10:eaam8460
Published by AAAS