Semaphorin-3 Promotes Specific Immunotherapy Effects on Experimental Food Allergy

Figure 3

Assessment of the role of Pak1 in RAS activation in sensitized mast cells. (a–c) Sensitized (Sens) and naïve control (Con) BMMCs were analyzed by RAS-specific ELISA. Violin plots show RAS.GTP levels (a), RAS.GDP levels (b), and RAS activation status (c). (d) Positive correlation between RAS activation and Pak1 mRNA levels in sensitized BMMCs. (e, f) Pak1 mRNA (e) and protein (f) levels in sensitized BMMCs. FTS: a pan inhibitor of RAS in the culture (75 μM). (g) A triple complex of Pak1, Sos1, and kRAS in sensitized BMMCs. (h–j) RAS activation in BMMCs after the treatment denoted on the axis. Pak1.OverEx: BMMCs with Pak1 overexpression. Control: BMMCs were transfected with empty plasmids. Sos1.RNAi (cRNAi): BMMCs were treated with Sos1 RNAi (or control RNAi). The data in the violin plots and boxplots are presented as median (IQR). Each bubble presents data obtained from one experiment. 1 [Student test (a–c) or ANOVA+Dunnett’s test (e, h–j)], compared with the saline group. Pearson correlation coefficient test was performed in (d).

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