Effect of MSP on RSK2 phosphorylation and its nuclear translocation: M-RON cells (3 × 106 cells per dish) in DMEM with 1% FBS were stimulated with MSP (2 nM), TGF-β1 (5 ng/ml), or MSP plus TGF-β1 for various times. Cellular proteins (50 μg/sample) from cell lysates were subjected to Western blot analysis using antibodies specific to regular or phospho-RSK (Ser380) and Erk1/2. A) MSP induced RSK2 phosphorylation and nuclear translocation. B) Effect of MSP and TGF-β1 on RSK2 phosphorylation and nuclear translocation. C) Immunofluorescent analysis of MSP or MSP plus TGF-β1-induced RSK2 nuclear translocation. M-RON cells were stimulated with MSP or MSP plus TGF-β1v for various times. Cells were fixed with cold acetone, blocked with 1% BSA, followed by incubation with antibody specific to RSK2. FITC coupled rabbit anti-mouse IgG was used as the detecting antibody. Immunofluorescence was observed by Olympus BT71 microscope equipped with fluorescent apparatus as previously described . Scale bars represent 5 μm.