Fig. 2

Ras isoforms differentially modulate reciprocal CD40 signaling. a Immunoblot analysis for checking the specificity of siRNA used for silencing respective isoforms. Only the Isoform-specific siRNA but not the control siRNA silenced the Ras isoforms. b Control siRNA treated P388D1 cells were stimulated with anti-CD40 antibody (3 μg/ml) for 15 min; using Western blot CD40 (3 μg/ml)-induced p38MAPK or ERK-1/2 activation was assessed. c-e siRNA treated P388D1 cells were stimulated with anti-CD40 antibody (3 μg/ml) for 15 min. siRNAs specific for H-Ras (c), K-Ras (d) and N-Ras (e) modulated the CD40 (3 μg/ml)-induced p38MAPK or ERK-1/2 activation, as assessed by Western blot. e-g siRNAs for H-Ras (f), K-Ras (g) and N-Ras (h) modulated the anti-CD40 antibody (3 μg/ml)-induced IL-12 or IL-10 productions as measured by using specific cytokine ELISA kits, in a macrophage-like mouse cell line P388D1. siRNA treated P388D1 cells were stimulated with anti-CD40 antibody (3 μg/ml) for 48 h. Data represented as mean ± SEM; *, p ≤ 0.05; ** p ≤ 0.005 compared with anti-CD40 stimulated P388D1 cells.