Figure 5.

Procaspase 3 protein expression. CuONP activates cell apoptosis pathway by auto-cleaving procaspase 3 into active caspase 3. We incubated CuONP of 100 μM with three lines of cells: SH-SY5Y (A), H4 (B), and PC12 (C). Cells were plated in 60 mm dish and treated with CuONP for 6 h in H4 and PC12 cells, and 16 h in SH-SY5Y cells. Cells were subjected to lyses in a RIPA buffer. The same amount of protein was run in 4%–12% Tris-Bis gel electro-transferred to nitrocellulose membrane. After being washed with PBST, the membranes were incubated with the first antibody 1:200 overnight and the secondary antibody 1:5000 for 2 h and developed a film by detection kit. The membranes were stripped by stripped buffer and re-incubated with first antibody actin to control the protein amounts. The films were scanned, and densities were analyzed by NIH ImageJ software. ** p < 0.01.