Figure 2
Intracellular calcium is involved in resistin-induced phosphorylation of c-Src.
(A) MDA-MB-231 cells were pre-incubated in Fluo-3 AM (5 μM) for 45 min. Calcium response was measured by confocal microscopy, where calcium concentration correlated with fluorescent intensity. (B) Cells were stimulated for 60 min at several concentrations of resistin. Western blots were performed on cell lysates (30 μg) and the membrane was probed for phospho-c-Src with c-Src as the control. (C) Cells were incubated with 25 ng/ml resistin for the indicated times. Western blots were conducted on cell lysates and the membrane was probed for phospho-c-Src with c-Src as the control. (D) After being serum-starved overnight, cells were treated with the indicated concentrations of resistin for 30 min. Western blots were conducted on cell lysates and the membrane was probed for phospho-c-Src with c-Src as the control. (E) Cells were incubated for 30 min with 25 ng/ml resistin in the presence of BAPTA-AM. Western blots were conducted on cell lysates and the membrane was probed for phospho-c-Src with c-Src as the control (F) Each column represents the average of 4 replicates of the Matrigel invasion assays. The results are displayed as mean ± SD from 3 independent experiments. *P < 0.05 vs. untreated condition.
