Fig. 2

AZ and/or MS-275 treatments increased cell cycle arrest and apoptosis of NB cells. a-b show propidium iodide analysis of cell cycle at 48 h after treatment with AZ(40 μM), MS-275(1.5 μM) and AZ + MS-275 (40 μM + 0.75 μM) in NB SH-SY5Y, indicating increase entry of SH-SY5Y cells into SubG0-phase (0.6%, %57 and %61; p ≤ 0.001) with decrease into S-phase (13%, 9% and 4%; p ≤ 0.05) and G2/M-phase (6%, 2% and 3%; p ≤ 0.05). c-d show western blot analysis of cell cycle inhibitor, p21, indicates 48 h treatment of MS-275 and AZ + MS-275 treatments cause of 4.5 and 5.5 induction of p21 (p < 0.01) of SH-SY5Y cells compare to control, respectively. e-f show in-cell western (0.75 μM and 1.5 μM MS-275 treatment; 48 h) analyses of proteins associated with cell cycle arrest and apoptosis in NB SH-SY5Y cells. Results indicate that levels of cyclin D1 (0.666 ± 0.010%; p = 0.0006), CDK4 (0.690 ± 0.033%; p = 0.0002) and BCL2 decreased significantly (0.376 ± 0.014%; p = 0.0035) while p16 CDK inhibitor significantly increased (2.528 ± 0.101%; p = 0.0002). g-h show western blot data using lysates from cells treated 48 h with 0.75 μM and 1.5 μM MS-275. p21 and p27 showed constant levels following treatment whereas cyclin D1 and CDK4 were reduced in expression. Expression of Ki67 proliferative marker decreased as well