Fig. 3
STAT6 negatively regulates HIF-1α protein synthesis. a, b U87MG cells transfected with non-targeting control siRNA (CONsi) or pooled siRNA targeting STAT6 (STAT6si) were exposed to 0% O2 for 18 h, followed by quantification of HIF-1α mRNA by RT-qPCR (a) and immunoblotting for HIF-1α and STAT6 (b). c Immunoblot of HIF-1α in CONsi or STAT6si-transfected U87MG cells exposed to 0% O2 for 18 h, followed by treatment with or without CHX (25 μM) for the indicated times (left) and summary data (right) showing actin-normalized HIF-1α levels, expressed as a percentage relative to time 0 (as 100%). Results are presented as means ± SD (error bars) of three independent experiments. d Immunoblot of newly synthesized HIF-1α in AHA-labeled U87MG cells exposed to 18 h of 0% O2. e, f U373MG cells transfected with empty vector (MOCK) or Myc-tagged full-length STAT6 (STAT6) were exposed to 0% O2 for 18 h, followed by quantification of HIF-1α mRNA by RT-qPCR (e) and immunoblotting for HIF-1α and STAT6 (f). g IF staining of HIF-1α in STAT6-transfected U373MG cells exposed to 0% O2 for 18 h. h Immunoblot of HIF-1α in MOCK- or STAT6-transfected U373MG cells treated as in c (left) and summary data (right) showing actin-normalized HIF-1α levels, expressed as a percentage relative to time 0 (as 100%). Results are presented as means ± SD (error bars) of three independent experiments. i Immunoblot of newly synthesized HIF-1α in AHA-labeled U373MG cells exposed to 18 h of 0% O2