Fig. 5.

Combined inhibition of PI3Kβ and MLK3 suppressed cell migration and invasion, and blocked the formation of lamellipodia and focal adhesions. a, b Boyden chamber migration assay in U-87 MG and U-118 MG cells treated with AZD6482 and URMC-099 for 6 h. c, d Boyden chamber invasion assay in U-87 MG and U-118 MG cells treated with AZD6482 and URMC-099 for 24 h. P values were determined by One-way ANOVA and Post Hoc multiple comparison Tukey HSD test. Compared with DMSO, *: p < 0.05; ***: p < 0.001; compared with AZD6482, ^a: p < 0.05; compared with URMC-099, ^b: p < 0.05. e Immunofluoresescence of U-118 MG cells after treatment with AZD6482 (30 μM) and URMC-099 (3 μM) for 3 h. Representative photographs show the effects of AZD6482 and URMC-099 on the formation of lamellipodia protrusions (white arrows) and FAs (yellow arrows). Bar = 20 μm. f Number of U-118 MG cells with lamellipodia was decreased by the combination of the combination of AZD6482 and URMC-099. g Number of FAs per U-118 MG cell was reduced by the combination of the combination of AZD6482 and URMC-099. P values were determined by One-way ANOVA and Post Hoc multiple comparison Tukey HSD test. *: p < 0.05; **: p < 0.01; ***: p < 0.001. (H) Immunoblotting of U-118 MG cells treated with AZD6482 (30 μM) and URMC-099 (3 μM) alone or in combination for 24 h. Expression of GAPDH was served as a loading control. Data were representative of two independent experiments