Akt is important for Sema
4D expression in PaTu8988 cells, inhibited by SAHA
. PaTu8988 cells were stimulated with indicated concentration of SAHA, cells were further cultured for indicated time, expressions of p-Akt (Ser 473), Akt, p-mTOR (Ser 2448), mTOR, p-S6K1 (Thr 389), S6K1, p-S6 (Ser 235/236), S6, SHP-2, EGFR, PDGFRβ and β-actin were detected by western blots (A-C). PaTu8988 cells were treated with Akt inhibitor perifosine (10 μM), mTORC1 inhibitor rapamycin (100 nM) or Erk/MAPK inhibitor PD98059 (10 μM), cells were further cultured for 24 h, expressions of p-Akt (Ser 473), Akt, p-S6 (Ser 235/236), S6, p-Gsk3β, β-actin, Erk1/2, p-Erk1/2 and Sema-4D were tested by western blots. Akt/mTOR/S6K1 phosphorylation in (A), Akt/S6 phosphorylation in (B), EGFR/PDGFRβ expression in (C), and Sema-4D expression in (D) were quantified after normalization to corresponding loading controls, and the number was presented as fold change vs. untreated ("C") control group. Experiments in this figure were repeated three times, and similar results were obtained.