PKA was not required for the rescue effect of A2A-R
Cells were transfected with an empty vector or a construct encoding p53-R175H or/and a construct encoding PKI along with a GFP vector as indicated. One day after transfection, cells were pretreated with or without H89 (10 μM) for 30 min before the addition of NGF (100 ng/ml), NGF plus CGS (1 μM), or NGF plus FK (10 μM) for 3 days. NGF-evoked neurite-bearing cells were quantified. Data represent the mean ± S.E.M. values from at least three independent experiments.
| Cell | CGS | FK | Inhibitor | Neurite-Bearing Cells | |
|---|---|---|---|---|---|
| Vector | R175H | ||||
| % | |||||
| PC-12 | − | − | − | 35.4 ± 5.2 | 14.3 ± 3.2a |
| PC-12 | + | − | − | 42.2 ± 4.8 | 42.7 ± 6.5b |
| PC-12 | − | + | − | 39.7 ± 1.3 | 42.8 ± 9.8c |
| PC-12 | − | − | PKI | 34.6 ± 3.4 | 14.2 ± 3.6a |
| PC-12 | + | − | PKI | 38.4 ± 1.8 | 39.0 ± 2.7b |
| PC-12 | − | + | PKI | 38.8 ± 8.1 | 15.6 ± 3.8a |
| PC-12 | − | − | H89 | 37.8 ± 4.9 | 11.4 ± 1.5a |
| PC-12 | + | − | H89 | 46.2 ± 0.8 | 44.4 ± 4.7b |
| A123 | − | − | − | 43.7 ± 1.5 | 22.7 ± 1.6a |
| A123 | + | − | − | 45.4 ± 4.0 | 45.1 ± 1.9b |
+, with the indicated drug; −, without the indicated drug.
↵ a Specific comparison between cells transfected with the indicated plasmid or a control vector (p < 0.001; two-way ANOVA).
↵ b Specific comparison between cells treated with or without CGS (p < 0.001; two way ANOVA).
↵ c Specific comparison between cells treated with or without FK (P < 0.001; two-way ANOVA).