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. Author manuscript; available in PMC: 2021 May 26.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2021 Apr 8;41(6):1987–2005. doi: 10.1161/ATVBAHA.121.316153

Fig. 2. The involvement of Grb2, PI3K, or SHP2 in Gab2-mediated inflammatory signaling.

Fig. 2.

(A) HUVECs were transfected with 200 nM scrambled, Grb2, SHP2, or Gab2 siRNA for 48 hours. The gene silencing was confirmed by immunoblotting. (B) The transfected cells were treated with TNFα, IL-1β (10 ng/ml) or LPS (500 ng/ml) for 6 h. VCAM1 expression was analyzed by immunoblot analysis. Band intensities were quantified by densitometry, and the quantified data were shown in the right-side panels. (C-H) HUVECs infected with control (C RV) or Gab2 retrovirus (Gab2 RV) were treated with LY294002 (5 μM) or SHP099 (0.5 μM) for 1 h. Then, the cells were treated with TNFα (C, D), IL-1β (E, F), or LPS (G, H) for 6 h. The total RNA was extracted from the cells and MCP1 (C, E, G), IL-8 (D,F,H) mRNA expression levels were measured by qRT-PCR. Results were expressed as relative expression to the control. One-way analysis of variance (ANOVA) was used to compare the data of experimental groups, Tukey’s post hoc multiple comparison test was used to obtain statistical significance.***, p<0.001, **, p<0.01; *, p<0.05; ns, no statistically significant difference***, p<0.001; ns, no statistically significant difference.