FIG. 4.

Knockdown of SOCS3 expression restores STAT3 phosphorylation in IL-6KO fibroblasts. IL-6KO dermal fibroblasts were grown as described and electroporated with SOCS specific siRNA and a scrambled control. Twenty-four hours post transfection, growth media was replaced with serum free media and treated up to 20 ng/mL rmIL-6 for 30 min. Total mRNA and protein were extracted from fibroblasts as described. Expression of SOCS3 (A) was determined via Real-time PCR utilizing an ABI 7000 SDS system and accompanying software. Phospho-STAT3 and total STAT3 was determined by Western blot (representative B) and differences determined by digital image analysis utilizing Image J (C). Data are expressed as a percent of untreated non-transfected control cells (A, C), mean±SE, (n=4, *Significantly different from 0 ng/mL, P≤0.05). SOCS, suppressor of cytokine signaling.