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. 2017 Dec 20;9(12):10294–10306. doi: 10.18632/oncotarget.23516

Figure 4. Inhibition of CDK5 signaling in fibroblasts abrogates TGF-β-induced responses.

Figure 4

(A) Fibroblasts transfected with indicated siRNA for 24 h followed by incubation with TGF-β2 (10 ng/ml) for further 24 h. Left panels, Western analysis. Representative images. Right panel, real-time qPCR. Results, normalized with GAPDH, are mean ± SD of triplicate determinations from an experiment representative of three. *p < 0.05. (B) Fibroblasts with CDK5 siRNA were transiently transfected with [SBE]4-luc, followed by TGF-β (10 ng/ml) for 24 h. Whole cell lysates were assayed for their luciferase activities. Results, normalized with renilla luciferase, represent the means ± SD from triplicate experiments. *p < 0.05. (C) Fibroblasts were incubated in media with roscovitine (10 μM) and TGF-β for 30 min, and whole cell lysates were examined by Western analysis. Representative images. (D) Fibroblasts were incubated with roscovitine (10 μM) for 30 min followed by incubation with TGF-ß (10 ng/ml) for 24 h. Left panels, Western analysis. Representative images. Right panel, real-time qPCR. Results, normalized with GAPDH, are mean ± SD of triplicate determinations from an experiment representative of three. * p < 0.05. (E) Fibroblasts were preincubated with TGF-ß for 24 h, followed by roscovitine (10 μM) for 24 h. Real-time qPCR results, normalized with GAPDH, are mean ± SD of triplicate determinations from an experiment representative of three. *p < 0.05.