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. 2011 Apr;178(4):1591–1604. doi: 10.1016/j.ajpath.2010.12.020

Figure 3.

Figure 3

The PTP1B−/− hepatocytes are hypersensitive to TNF-α– and IL-6–mediated signaling. A: Left: Primary hepatocytes (PTP1B+/+ and PTP1B−/−) were stimulated with 10-ng/mL TNF-α for the indicated periods. Control cells were maintained in the absence of cytokines. At the end of the culture time, cells were lysed and 50 μg of total protein was analyzed by Western blot with antibodies against phospho-JNK (pJNK) (Thr183/Tyr185), JNK, phospho-c-Jun (Ser73), phospho-ERK (pERK) (Thr202/Tyr204), and ERK. Right: Immortalized PTP1B+/+ hepatocytes were transfected with PTP1B or scramble siRNA oligos. After 48 hours, cells were stimulated with 10-ng/mL TNF-α or left untreated. Total protein was analyzed by Western blot with the indicated antibodies. Results shown are representative of two independent experiments. B: Left: Primary hepatocytes were cultured as described in A and stimulated with 10-ng/mL IL-6 for the indicated periods. Control cells were maintained in the absence of cytokines. At the end of the culture time, cells were lysed and 50 μg of total protein was analyzed by Western blot with antibodies against phospho-STAT3 (pSTAT3) (Tyr705), STAT3, pERK (Thr202/Tyr204), and ERK. Right: Immortalized PTP1B+/+ hepatocytes were transfected with PTP1B or scramble siRNA oligos. After 48 hours, cells were stimulated with 10-ng/mL IL-6 or left untreated. Total protein was analyzed by Western blot with the indicated antibodies. Representative autoradiograms are shown. Similar results were obtained in two series of independent experiments.