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. 2012 Mar 16;287(19):15263–15274. doi: 10.1074/jbc.M111.303958

FIGURE 5.

FIGURE 5.

p65 knockdown in PTP1B−/− hepatocytes recovers the apoptotic response to TGF-β and Smad2 and Smad3 phosphorylation and nuclear translocation, coincident with lower Smad7 induction. PTP1B+/+ and PTP1B−/− cells were transfected with either an unspecific siRNA (Uns.) or p65 siRNA (p65) and then treated or not with TGF-β. A, percentage of apoptotic nuclei at 30 h. B, Western blot of total lysates at 2 h. C, percentage of p-Smad2-positive nuclei in at least 10 different fields for experiment (n = 3) at 2 h. D, Western blot of cytosolic/nuclear extracts of PTP1B−/− treated for 2 h with TGF-β. β-Actin was used as loading control for total, and cytosolic extracts and PCNA were used as loading control for nuclear extracts in B and D. Data represent the mean ± S.E. (error bars) of three independent experiments. Student's t test was calculated as unsilencing siRNA versus p65 siRNA for each experimental condition in each cell type: *, p < 0.05; **, p < 0.01; ***, p < 0.001.