Research Paper Volume 13, Issue 15 pp 19375—19396

Figure 8. Activation of AKT by THBS4/integrin α2 axis in HUVECs. (A, B) rTHBS4 induced the phosphorylation of AKT in HUVECs. n= three independent experiments. The HUVECs were treated with and without rTHBS4 for 1h and the whole cell lysates were analyzed by WB for levels of downstream kinases. (C, D) p-AKT activation occurred in a time-dependent manner in rTHBS4-treated cells. n= three independent experiments. (E, F) The HUVECs were treated with rTHBS4 for 1h in the presence of integrin α2-specific antibody or PI3K inhibitor LY294002. Western blot analysis showed that rTHBS4 increased Akt phosphorylation in HUVECs, while this effect was alleviated by blocking integrin α2 or inhibiting Akt. n= three independent experiments. (G) Transwell migration assay to investigate the effect of PI3K inhibition and integrin α2 blockade on HUVEC migration after treatment with rTHBS4. n= 3 wells per group. (H) In vitro tube formation to investigate the effect of PI3K inhibition and blocking integrin α2 on HUVEC tube formation. n= 3 wells per group. (I) Quantification of G. (J) Quantification of H. (K, L) The CM from BM-MSCs stimulated by H. Pylori was applied to HUVECs in the presence or absence of integrin α2-specific antibody or PI3K inhibitor LY294002 for Transwell migration assay and tube formation assay. n= 3 wells per group. (M) Quantification of K. (N) Quantification of L.