Fig. 3.

Combined inhibition of PI3Kβ and MLK3 exhibited synergistic inhibitory effect on glioblastoma cell proliferation. a Expression pattern of PI3K/Akt pathway molecules in a series of glioblastoma cell lines. Normal human astrocytes were used for comparison. The expression of β-actin was used as a loading control. b, c Cell proliferation assay in 7 GBM cell lines treated with AZD6482 or URMC-099 alone at different concentrations for 48 h. DMSO was used as a carrier control. d IC₅₀ of AZD6482 and URMC-099 in 7 GBM cell lines. e, f U-87 MG and U-118 MG cells were treated with AZD6482 and URMC-099 at fixed ratios (6:1 and 10:1, respectively) for 48 h. Their combination effect on GBM cell proliferation was shown. P values were determined by Two-way ANOVA and Post Hoc multiple comparison Tukey HSD test. Compared with AZD6482, *: p < 0.05; **: p < 0.01; ***: p < 0.001; compared with URMC-099, ^#: p < 0.05; ^##: p < 0.01; ^###: p < 0.001. g, h Fa-CI plots were automatically generated by Compusyn software according to the Chou-Talalay method, representing the CI values of combination treatment of AZD6482 and URMC-099. Two dashed lines indicated the CI values of 0.9 and 1.1