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. 2018 May 7;217(5):1701–1717. doi: 10.1083/jcb.201706118

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© 2018 Papadopoulos et al.

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Figure 9. — The Fer–TMF-1–Brg-1 axis regulates proliferation by controlling p21 protein levels. (a) Knockdown of Fer, TMF-1, and Brg-1 inhibit proliferation in response to PDGF-BB and serum. Proliferative response was evaluated by measuring incorporation of a fluorescent DNA-binding dye; fluorescence values were plotted against increasing concentrations of PDGF-BB. The statistical significance of the difference between control and knockdown cells at 20 ng/ml PDGF-BB was analyzed with two-tailed t test. Error bars indicate SD. *, P < 0.05; ***, P < 0.001. (b) The efficiencies of the knockdowns were determined by immunoblotting. Ib, immunoblotting. (c–f) Immunostaining of p21 protein expression upon Fer, TMF-1, and Brg-1 knockdown. Cells treated with control siRNA or siRNA for Fer, TMF-1, or Brg-1 were stained for p21 protein (green) and PDGFRβ (red); chromatin was stained with DAPI (blue). Bars, 10 µm. (g) The percentage of p21-expressing cells was counted using the automatic pipeline in Cell Profiler.