| Adherent | Suspension |
---|
v-Src Substrates | Similar levela
| More in FAK+/+ [v-Src] | More in FAK-/- [v-Src] | Similar level | More in FAK+/+ [v-Src] | More in FAK-/- [v-Src] |
---|
Vinculin (poY100) |  | 5 – 6 |  |  | 7 – 8 |  |
p190RhoGAP (poY)* |  |  | 1.5 – 2 |  |  | 1.5 – 2 |
Shc (poY239/poY240) | Yes | Â | Â | Yes | Â | Â |
Sam68 (poY)* | Â | Â | 2 | Yes | Â | Â |
PLC-γ1 (poY783) | Yes |  |  | Yes |  |  |
PKC-δ (poY311) |  |  | 2 | Yes |  |  |
Paxillin (poY118) | Â | Â | 4 | Â | Â | 10 |
Paxillin (poY31) | Yes |  |  |  | 2 – 3 |  |
Eps8 (poY)*, b
| Yes | Â | Â | Yes | Â | Â |
p56 Dok-2 (poY351) | Â | 5 | Â | Â | Â | 2 |
Crk (poY)* | Â | Â | 2 | Â | Â | 2 |
Cortactin (poY421)c
| Yes | Â | Â | Â | 2 | Â |
Connexin43 (poY265)d
|  |  | 2 |  | 2–3 |  |
p120Catenin (poY228) | Â | 2 | Â | Â | Â | 2 |
p130 CAS (poY410) | Â | 2 | Â | Â | 2 | Â |
Annexin II (poY)* | Yes | Â | Â | Yes | Â | [3]e
|
- *blotting with mAb 4G10 anti-PTyr was used to ascertain the level of tyrosine phosphorylation after immunoprecipitation with substrate-specific Abs.
-
astatistically equal level of relative substrate phosphorylation between FAK+/+[Src] and FAK-/-[Src] cells, based on triplicate experiments. Relative phosphorylation for a given experiment is calculated as the phospho- substrate signal normalized to total substrate protein level, then normalized to loading control proteins. Note that Src transformation increases GAPDH protein levels ~2-fold and decreases actin protein level ~2-fold in the FAK+/+ background in adherent cells only.
-
bfor both 97 kDa and 68 kDa Eps8 isoforms.
-
cin adherent cultures, the 80 kDa cortactin isoform protein level is reduced in the absence of FAK, but the relative phosphorylation levels are similar.
-
dconnexin-43 (poY265) levels were not normalized.
-
erelative decrease in phospho-annexin II levels in suspended FAK-/-[v-Src] cells.