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. 2020 Mar 17;22(10):1452–1462. doi: 10.1093/neuonc/noaa064

Fig. 4.

Fig. 4

C5a increases ZEB1 expression via C5aR1-mediated p38 MAPK activation. (A) Invasion of GSC11 GBM cells co-cultured with MSLC09-03 cells in transwells after transfection with a C5aR1-siRNA (A) or treatment with the C5aR1 antagonist W54011 (B) as indicated (n = 3). (C) RT-qPCR for ZEB1 levels in X01 cells treated with the C5aR1 antagonist W54011 in co-culture with MSLC09-03 (n = 3). (D) Invasion/migration with GBM cells transfected with siRNA as indicated and co-cultured with MSLC09-03. (E) Infiltration of GBM cells into a 3D collagen matrix premixed with MSLC09-03 after transfection with a p38-siRNA. The inset shows spheroids at the initial time. (F) Western blot for p-p38 in X01 co-cultured with MSLC09-03 in the presence of an anti-C5a antibody (n = 3). (G) Western blot for p-p38 and ZEB1 in X01 treated with the C5aR1 antagonist W54011 in co-culture with MSLC09-02 (n = 3). (H) RT-qPCR for ZEB1 in X01 co-cultured with MSLC09-02 after siRNA transfection (n = 3). (I–K) Kaplan–Meier survival of human all brain tumors from REMBRANDT (I), TCGA GBM (J), and mesenchymal type of TCGA GBM (K) patients with high or low C5aR1 median expression. *P < 0.05, **P < 0.01, ***P < 0.001.