Fig. 2

RAGE inhibition hampers Ins-induced signaling in BC cells. Representative immunoblots showing the phosphorylation of IR (Y1135/1136) (A, F, K, O), IRS1 (Y612) (B, G, L, P) and AKT (S473) C, H, M, Q in MCF-7 and ZR75 cells treated with Ins (20 nM, 15 min) alone and in combination with FPS-ZM1 (2 μM and 10 μM, 24 h) or RAP (50 μM, 24 h), as indicated. FPS-ZM1 (10 μM, 24 h) and RAP (50 μM, 24 h) interfere with the upregulation of CD1 mRNA (D, I) and protein expression E, J, N, R in MCF-7 and ZR75 cells stimulated with Ins (20 nM, 4 h), as indicated. Total proteins and β-actin serve as loading control. In qRT-PCR experiments, values are normalized to the 36B4 gene expression and shown as fold change of CD1 mRNA expression induced by Ins compared to cells treated with vehicle (−). Data shown are mean ± SEM of at least three independent experiments performed in duplicate. (*) p < 0.05; (***) p < 0.001