SCF reduces hypoxia induced IM-PTEC apoptosis and induces phosphorylation of Bad.

<p>(A) SCF reduces caspase 3 activity following hypoxia. IM-PTEC cells were subjected to in vitro hypoxia and cultured with 0 ng or 100 ng SCF/ml medium for 24 hours. Cell lysates were collected and assayed for caspase 3 activity. To correct for equal input, values are expressed as activity per µg protein. SCF reduces caspase 3 activity in IM-PTEC cells following in vitro hypoxia (*<i>P</i> = 0.026). Data are expressed as mean±SEM. Results are obtained from 2 separate experiments with 6 measurements each. (B) IM-PTEC cells were serum starved and exposed to 0, 10 or 100 ng SCF/ml for 5, 10 and 15 minutes. Protein samples were analyzed using Western blot. SCF induced phosphorylation of Tyr719 of c-KIT; β actin was used as loading control. (C) SCF results in phosphorylation of Ser473 of Akt; total Akt expression was used as loading control (upper panel); SCF induces phosphorylation of Ser136 of Bad; β actin was used as loading control (lower panel). Normalized densitometric analyses of the immunoblots are presented as the ratio phosphorylated protein/loading control. Data shown here are representative results obtained from 3 separate experiments. (D) Renal tissue samples obtained from sham and animals subjected to ischemia were analyzed for phosphorylation of Akt (p-Ser473 Akt) and c-KIT (p-Tyr719 c-KIT). Expression of total Akt and c-KIT were used as loading control.