Fig. 4.

The effect of rotenone on the expression of apoptotic proteins BCL2, BAX, CASP9, PARP, and cleaved CASP3 in SH-SY5Y cells. (A) The effect of rotenone on the expression of BCL2, BAX, CASP9, PARP, and cleaved CASP3 by Western blot analysis. Cells were treated with 10 µM rotenone for 24 h. (B) Western blot assay to determine the effect of naringin on rotenone-induced expression of the apoptotic proteins, BCL2, BAX, cleaved CASP9, cleaved PARP, and cleaved CASP3 in SH-SY5Y cells. Cells were pretreated with 10 µM rotenone for 24 h, with 10 µM naringin added 4 h prior to rotenone treatment. β-Actin was detected as an internal standard protein. (C) Effect of naringin on rotenone-induced activity of CASP3 in SH-SY5Y cells. Cells were pretreated with 100 µM rotenone for 24 h and 10 µM naringin was then added 4 h prior to rotenone treatment. The cleaved CASP3 substrate acetyl-Asp-Glu-Val-Asp-p-Nitroanilide (Ac-DEVD-p-NA) was measured at 405 nm. CASP3 was used as a positive control. The CASP3 inhibitor (Ac-DEVD-CHO) added with CASP3 was used as negative control. Values are presented as mean±SEM. Three independent experiments were performed for this assay. ^*p<0.05 compared to non-treated cells; ^†p<0.05 compared to rotenone-treated cells.