Fig. 5.

p38MAPK contributes to FKBP9’s oncogenic function in GBM cells. a Cell lysates from SF-539 and T98G cells treated as in Fig. 2b were analyzed by IB for key protein levels of MAPK pathway. b Protein levels of p38 and its downstream genes in SF-539-FKBP9-WT/FKBP9-M541I and T98G-FKBP9-WT/FKBP9-M541I cells were detected by IB. c, d SF-539-FKBP9-WT and T98G-FKBP9-WT cells were treated with vehicle or 5 μM SB201290/SB203580 and the ability of these cells to form colonies and spheres was determined. e Nestin and Sox2 levels of SF-539 and U-87 MG cells spheres were detected by IB. f, g SF-539-PCDH-vector and FKBP9-WT cells were transfected with two siRNA duplexes targeting p38 (sip38) or control siRNA (siCtrl) for 48 h. Colony formation and sphere formation assays were performed. Nestin and Sox2 expression were detected by IB. h Protein expression of p38’s upstream regulators in SF-539-FKBP9-WT and T98G-FKBP9-WT cells was detected by IB. i SF-539-FKBP9-WT and T98G-FKBP9-WT cells were treated with vehicle (DMSO), AZD9291 (1 μM), BAY-11-7086 (10 μM), ICG-001 (5 μM), LY364947 (10 μM), MK2206 (2 μM), PD98059 (10 μM), NQDI-1 (5 μM), Rapamycin (2 μM), SB203580 (2 μM), Takinib (5 μM), XAV939 (10 μM) or 4PBA(50 μM), respectively. IB analysis was performed for p-p38, p38 and pHSP27. GAPDH was used as a loading control. j Cells transfected with PCDH-vector or FKBP9-WT were treated with vehicle or 5 μM NQDI-1, or transfected with two siRNA duplexes targeting ASK1 (siASK1) or control siRNA (siCtrl) for 48 h. Colony formation assays were performed. All experiments in this figure were performed three times with comparable results. Data are represented as mean ± S.D. (**p < 0.01, ***p < 0.001)