Figure 4.

SULF2-mediated up-regulation of STAT3 target genes depends on both GLI1 and STAT3. A, assessing the role of STAT3 in regulating SOCS2, SOCS3, PIM1, and FLT3 expression in Huh-7 cells by qPCR. Cells were transduced with either Adv-Null (Control, n = 3) or Adv-SULF2 (SULF2, n = 3) and transfected with either nontargeting siRNA (n = 3) or siRNA against STAT3 (siSTAT3, n = 3). Results are expressed as means ± S.E. (error bars). B, promoter diagrams are reproduced for SOCS2, SOCS3, PIM1, and FLT3. Relative position of putative STAT3 consensus binding sites are indicated in blue. ChIP-PCR experiments are expressed as STAT3 -fold enrichment at these binding sites in Huh-7 cells under conditions of Adv-Null (n = 3) and Adv-SULF2 (n = 3) transduction. Results are expressed as means ± S.E. C, representative co-immunoprecipitation assays in Huh-7 cells after co-transfection of STAT3-FLAG and GLI-His constructs under conditions of transduction with Adv-Null (left) and Adv-SULF2 (right) transduction. MW, molecular weight presented in kilodaltons.