Fig. 6

Sema5A promotes the activation of AKT-mTOR pathway in CD4 + T cells through PlexinA1. A. Activated CD4⁺ T cells were cultured for 24 h in medium containing Sema5A or PBS solution, followed by stimulating for 1 h with the Th17 cocktail; the phosphorylation of AKT and mTOR was detected by Western-blotting. B. Activated CD4⁺ T cells were transfected with siRNA-Plexin A1- or universal control siRNA (NC) for 24 h, and resuspended in fresh medium containing Sema5A or PBS solution for another 24 h, followed by stimulating for 1 h with the Th17 cocktail; the phosphorylation of AKT and mTOR was detected by Western-blotting. * : P < 0.05, ** : P < 0.01. C. Activated CD4⁺ T cells were cultured for 4 h in medium containing pictilisib (PI3K inhibitor), MK-2206 (AKT inhibitor), and temsirolimus (mTOR inhibitor), then cells were resuspended in medium with Sema5A or PBS solution for 24 h under Th17-inducing conditions, and the IL-17 A level in the culture supernatant was detected by ELISA. Tukey test *: P < 0.05