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Fig. 3 | BMC Musculoskeletal Disorders

Fig. 3

From: Upregulation of TGF-β-induced HSP27 by HSP90 inhibitors in osteoblasts

Fig. 3

HSP90 inhibitors, geldanamycin, onalespib and 17-DMAG do not affect the phosphorylation of Smad2 induced by TGF-β in osteoblastic MC3T3-E1 cells. The cultured osteoblasts were pretreated with 0.3, 0.7 or 1.0 μM of geldanamycin (A), 1 μM of onalespib (B), 0.3, 0.7 or 1.0 μM of 17-DMAG (C) or vehicle for 60 min, and then incubated by 3 ng/ml of TGF-β or vehicle for 60 min. Cell extracts were analyzed by SDS-PAGE and Western blotting using antibodies of phospho-specific Smad2 or Smad2. The histogram shows the quantitative representations of phosphorylated Smad2 normalized with each total Smad2 gained from laser densitometric analysis. The levels were expressed as the fold increase to the basal levels presented as lane 1. A, B Triplicate determinations of Western blot analysis were performed corresponding to three independent cell preparations. Each value represents the mean ± S.E.M. of triplicate determinations from three independent cell preparations. *P < 0.05, compared to the value of the control cells without TGF-β-stimulation. N.S. means no significant difference between the indicated pairs

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