Figure 4
Hypoxia induces MIF mRNA (A) and protein levels (B, C) via HIF-1α pathway in HUASMCs. Growth-arrested HUASMCs were transfected with wild type HIF-1α, HIF-1α-siRNA or scrambled siRNA expressing plasmids for 24 h and then exposed to normoxia or hypoxia (3% O2) for 24 h. (A) Real-time PCR results. MIF mRNA expression were assayed by Q-PCR (n = 3 in each group). * P < 0.05 vs control cells under normoxia. # P < 0.05 vs control or scrambled siRNA transfection under hypoxia. (B) ELISA results. MIF protein released into cell culture media was measured by ELISA (n = 3 in each group). * indicates P < 0.05 vs control cells under normoxia. # P < 0.05 vs control or scrambled siRNA transfection under hypoxia. (C) Western blot results. Representative Western blot (top) and values of total MIF production (mean ± SEM of 3 experiments, bottom). Results of total MIF protein production were obtained from densitometric analysis and expressed as ratio of MIF/β-actin. * indicates P < 0.05 vs control cells under normoxia. # P < 0.05 vs control or scrambled-siRNA transfection under hypoxia. HIF-1: wild type HIF-1α plasmid transfection; Scrambled-siRNA: scrambled-siRNA plasmid transfection; HIF-1α-siRNA: HIF-1α-siRNA plasmid transfection.