FIGURE 7.
CCL2 protects PC3 cells from autophagic death. A, synchronized PC3 cells were plated in serum-free medium for 24 h prior to treatment with increasing concentrations of rapamycin (0-40 nm) with or without CCL2 (100 ng/ml). Cell viability was assessed by WST-1 dye conversion 24 h post-treatment. a, the survival curves represent the average absorbences (440 nm) of n = 5 samples for each condition with or without CCL2 (100 ng/ml), in response to increased rapamycin concentration. b, the data represented in a were normalized to 0 nm rapamycin for each condition (with or without CCL2) and are expressed as relative percentage of survival. B, a subset of cells treated as described in a with or without 20 nm rapamycin were analyzed by immunoblotting using survivin antibody. C, the effect of Akti-X on phospho-p70S6 kinase (Thr389) in CCL2-treated cells at 48 and 72 h post-treatment by Western blot analysis as compared with LC3B and survivin protein levels. D, IGF-1 (100 ng/ml) induces p70S6K phosphorylation (Thr389) only at early time points in PC3 cells as determined by Western blot analysis of cell lysates, as indicated. β-Actin was used as a loading control.