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

Figure 2. TGF-β stimulates p35 expression and CDK5 activation.

Figure 2

Confluent human foreskin (A–E) or mouse embryonic (C) fibroblasts were incubated in media with TGF-β (10 ng/ml). (A) Real-time qPCR. Results, normalized to GAPDH, represent means ± SD of triplicate determinations from an experiment representative of three. *p < 0.05. (B) Cultures were pre-incubated with U0126 (10 μM) or SB431542 (5 μM) for 30 min followed by TGF-β incubation for 24 h. Left panel, real-time qPCR. Results, normalized to GAPDH, represent means ± SD of triplicate determinations from an experiment representative of three. *p < 0.05. Right panels, Western analysis, representative images. (C) Smad3-null and wildtype MEFs were incubated with TGF-β for 4 h. Results of qPCR, normalized to GAPDH, represent means ± SD of triplicate determinations from an experiment representative of three. *p < 0.05. (D) Foreskin fibroblasts were incubated with TGF-β for 60 min, and whole cell lysates were subjected to Western analysis. Representative images. Band intensities normalized to tubulin shown below. (E) Foreskin fibroblasts were incubated with TNF-α, PDGF or TGF-β for 60 min, whole cell lysates were immunoprecipitated with CDK5 antibodies, incubated with Histone H1 and phosphorylation examined using p-Histone H1 antibody. Band intensities normalized to input tubulin are shown below.