IFN-у /STAT1 signaling is downregulated in cells with active Hh/GLI signaling.

<p><b>A</b>) The effect of SOCS1 expression on IFN-у/STAT1 signaling was analyzed in HaCaT cells expressing FLAG tagged SOCS1 (pLL-SOCS1) or empty viral vector as control (pLL). 48h hour post transfection cell were treated with 1 ng/ml IFN-у for 6h and assayed for mRNA levels of known IFN-у target genes by qRT-PCR. Fold change refers to the ratio of IFN-у treated to untreated samples. <b>B</b>) qRT-PCR of GLI2act-HaCaT cells expressing GLI2 (+DOX) for 72h and subsequently treated with 1 ng/ml IFN-у for 6h. mRNA levels IFN-у target genes (HLA-DRA, ICAM1, IFIT1, and p21) are shown as ratios to untreated control. PTCH was used as marker for GLI2 activity. <b>C</b>) Western blot showing STAT1 phosphorylation in GLI2act-HaCaT cells treated with doxycycline (DOX) for 72h followed by a 2h treatment with IFN-у. <b>D</b>) qRT-PCR of IFN-у target gene expression in DAOY cells treated with SAG or DMSO control for 144h followed by 4h of IFN-у treatment. mRNA levels are shown as ratio of treated to untreated samples. Data are given as mean ± SD of biological triplicates. <b>E</b>) STAT1 phosphorylation in SAG/IFN-у treated DAOY was analyzed by Western blot.