TABLE 3.
Effects of Src, Ras, PKC, and PI3K on survival and differentiation responses of PC12 cellsa
| Treatment | % Survival | % Differentiation |
|---|---|---|
| PC12 − NGF | 48 ± 4 | 0 |
| PC12 + NGF | 74 ± 6 | 60 ± 6 |
| Ras− − NGF | 49 ± 5 | 0 |
| Ras− + NGF | 48 ± 7 | 0 |
| Src− − NGF | 8 ± 2 | 0 |
| Src− + NGF | 29 ± 6 | 38 ± 6 |
| SrcO − NGF | 66 ± 4 | 0 |
| SrcO + NGF | 84 ± 8 | 75 ± 6 |
| LY, 50 μM | 12 ± 4 | 0 |
| LY + NGF | 48 ± 3 | 0 |
| W, 100 nM | 22 ± 7 | 0 |
| W + NGF | 52 ± 6 | 0 |
| PMA, 1 μM | 47 ± 5 | 0 |
| PMA + NGF | 85 ± 6 | 79 ± 8 |
| aPKC, 50 μM | 5 ± 1 | 0 |
| aPKC + NGF | 24 ± 3 | 30 ± 4 |
| cPKC, 50 μM | 23 ± 3 | 0 |
| cPKC + NGF | 59 ± 6 | 54 ± 5 |
| CH, 6 μM | 2 ± 1 | 0 |
| CH + NGF | 42 ± 6 | 32 ± 8 |
| PD, 60 μM | 60 ± 4 | 0 |
| PD + NGF | 79 ± 6 | 0 |
| SB, 20 μM | 69 ± 6 | 0 |
| SB + NGF | 73 ± 6 | 0 |
| Cu, 20 μM | 0 | 0 |
| Cu + NGF | 56 ± 6 | 37 ± 4 |
PC12 cells were plated and after 3 days washed five times with serum-free medium and cultured with or without NGF in a serum-free environment for 48 h; then all cells in the well were removed, stained with trypan blue, and counted. To score cells for differentiation, the cells were cultured with NGF for a total of 6 days and scored as described in Materials and Methods. The experiment was conducted using triplicate treatments and repeated twice. Cells were pretreated with LY294002 (LY), wortmannin (W), aPKC (myristoylated pseudosubstrate peptide), cPKC (myristoylated pseudosubstrate peptide), chelerythrine chloride (CH), PD98059 (PD), SB202190 (SB), or curcumin (Cu), for 1 h prior to addition of NGF. PMA was added for 48 h to remove cPKC and nPKC isoforms (56).