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. 2014 Mar 23;5(8):2131–2148. doi: 10.18632/oncotarget.1852

Fig 6. Jak1, but not Jak2, is required for STAT activation by IL-6 family cytokines.

Fig 6

(A) Jak2−/− MEFs were transiently transfected with a plasmid coding for eGFP or Jak2. Expression of Jak2 was assessed by Western blotting 48 h later. (B) Jak2−/− MEFs transiently transfected with eGFP or Jak2 were stimulated with increasing concentrations of Hyper-IL-6 (0-20 ng/ml) for 15 min. (C) Jak2−/− MEFs transiently transfected with Jak2 were stimulated with 10 ng/ml Hyper-IL-6, CT-1 or OSM for 15 min. Cells were pre-treated with 100 μM TBB for 90 min where indicated. (D, E) Jak1−/−, Jak2−/− and wildtype MEFs were stimulated with different concentrations (0-10 ng/ml) of Hyper-IL-6 or CT-1 for 15 min. Cells were pre-treated with 100 μM TBB for 90 min where indicated. (F) Jak1−/− MEFs were transiently transfected with a plasmid coding for eGFP or Jak1. Expression of Jak2 was assessed by Western blotting 48 h later. Lysate of wildtype MEFs served as positive control. (G) Jak1−/− MEFs transiently transfected with eGFP or Jak2 were stimulated with increasing concentrations of Hyper-IL-6 (0-20 ng/ml) for 15 min. Cells were pre-treated with 100 μM TBB for 90 min where indicated. Phosphorylation of STAT3 was determined in all experiments by Western blotting, and STAT3 as well as β-actin served as loading controls. Shown is one representative Western blot from at least three independent experiments.