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. 2011 Nov 17;6(11):e26674. doi: 10.1371/journal.pone.0026674

Figure 10. Feedback inhibition of Jak2/Stat5 pathway occurs via formation of a Jak2-IGFR-SOCS6 complex.

Figure 10

(A) PC12 cells stably transfected with SOCS6-EGFP or EGFP vector alone were stimulated with IGF-1 for 0 m, 5 m and 15 m (m = minute). Jak2 was immunoprecipitated and Western-blotted with anti-SOCS6 (top panel) or anti-Jak2 (bottom panel) antibodies. (B) PC12 cells transiently transfected with control or SOCS6-siRNA was stimulated with IGF-1 for 15 minutes. Jak2 was immunoprecipitated and Western-blot was performed using anti-IGFR antibody. The membrane was subsequently stripped and reprobed with anti-SOCS6 and anti-Jak2 antibodies. (C) PC12 cells stably transfected with SOCS6-EGFP or EGFP vector alone were stimulated with IGF-1 for 10 minutes. Stat5 was immunoprecipitated and Western-blot was performed using pY20 antibody. The membrane was subsequently stripped and reprobed with anti-Stat5 antibody. (D) PC12 cells transiently transfected with control or SOCS6-siRNA was stimulated with IGF-1 for 10 minutes. Stat5 was immunoprecipitated and Western-blot was performed using pY20 antibody. The membrane was subsequently stripped and reprobed with anti-Stat5 antibody. All the experiments were independently repeated 3 times with similar results.