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. 2019 Jul 30;33(11):11706–11720. doi: 10.1096/fj.201900698R

Figure 3.

Figure 3

Involvement of SHP2 in PAF-induced eosinophil migration. A) Immunofluorescent staining of CCR3 in rat peritoneal eosinophils. B) Enhancement of the phosphorylation of SHP2 and ERK1/2 by PAF. Rat peritoneal eosinophils were treated with PAF at 10 μM for the indicated times and harvested for preparation of protein lysates and Western blotting. C) The specificity of SHP2 inhibitors. The primary rat eosinophils were treated with PAF at 10 μM for 7 min following the pretreatment with either PHPS1 at 50 μM or RMC4550 at 1.0 μM for 1 h. Cells were then harvested for preparation of protein lysates and Western blotting. D, E) Attenuation of eosinophil migration by SHP2 inhibitors PHPS1 and RMC4550. Rat peritoneal eosinophil migration toward 10 μM PAF was analyzed by transwell inserts in the presence or absence of the indicated concentration of PHPS1 or RMC4550 for 3 h. The results were expressed as the number of migrated cells in bottom well. Data are presented as means ± sd (n = 3). ++P < 0.01 vs. without PAF, PHPS1 and RMC4550; **P < 0.01 vs. 10 μM PAF alone.