A–B, U251-MG cells were treated with BAY-11-7085 (BAY-11; 10 μM) for 2 h prior to JSI-124 (1 μM) treatment for 1 h. Isolation of RNA, generation of cDNA and quantitative RT-PCR was performed for IL-6 (A), IL-8 (A) and SOCS3 (B) expression. Data represent mean ± SD, replicates of three (*, p<0.01; **, p<0.001; Students t-test). The experiment was repeated, and similar results were observed. C–F, U251-TR/sh-p65 cells were treated with tetracycline (Tet) for 48 h to decrease p65 mRNA expression (C). After 48 h of Tet, cells were then treated with JSI-124 (1 μM) for 1 h. RNA was isolated, followed by generation of cDNA and quantitative RT-PCR was performed for IL-6 (D), IL-8 (E), and SOCS3 (F). Data represent mean ± SD, replicates of three (*, p<0.05; **, p<0.005; Students t-test).