Fig. 7
STAT6 expression is downregulated in GSCs. a Immunoblot of the indicated STATs in patient-derived GSCs (GSC1 and GSC2), non-tumor brain tissue (NTT), and high-grade glioma (HGG) tissue. STAT6 antibodies: #1, Cell Signaling Technology; #2, BD Biosciences; #3, Santa Cruz. b-d Immunoblot of STAT6 and STAT1 (b) and MSP (c) and bisulfite sequencing (d) of STAT6 promoter CpG islands in GSCs. e GSCs were treated with the indicated concentrations of 5-Aza for 72 h, followed by quantification of STAT6 mRNA by RT-qPCR (top) and immunoblotting for STAT6 and STAT3 (bottom). Results are presented as means ± SD (error bars) of three independent experiments (*p < 0.05). f Immunoblot of STAT6 and the indicated DNMTs in GSCs transfected with the indicated DNMT siRNA or control siRNA. g, h Immunoblot of HIF-1α and STAT6 in GSC1 cells transfected with empty vector (MOCK) or Myc-tagged full-length STAT6 (STAT6) and exposed to 21% or 0% O2 for 18 h (g) and IF staining of HIF-1α in STAT6-transfected GSC1 cells exposed to 0% O2 for 18 h (h). i CONsi- or STAT6si-transfected GSC1 cells were treated as in Fig. 6e, followed by immunoblotting for indicated proteins