FIG. 3.

The expression of EGR-1 is increased in dAT-MSCs. (A) nAT-MSCs and dAT-MSCs were cultured under normoxic (20% O₂) or hypoxic (5% O₂) conditions. The mRNA and protein levels of transcription factor EGR-1 were examined by a qRT-PCR and an immunoblot analysis, respectively, normalized to β-actin and Lamin B. (B) PTEN and GGPS1 expression was determined by a qRT-PCR and normalized to β-actin. (C) Insulin (1,000 nM) was added to the culture medium for 60 min to stimulate the phosphorylation of IRS-1 (P-IRS-1). P-IRS-1 at serine 636/639 and total IRS-1 (T-IRS-1) were analyzed by immunoblotting. (D) The mRNA expression level of interleukin-6 (IL-6) was assessed by a qRT-PCR and normalized to β-actin, and the quantitative protein concentration of IL-6 present in cell culture supernatants was measured by the IL-6 High Sensitivity Human ELISA Kit (D6050; R&D systems). (E, F) The mRNA expression levels were assessed by a qRT-PCR and normalized to β-actin to determine the expression of bFGF and TGF-β (E), Cyr61, Col4, and Inαv (F). White and black bars indicate normoxic and hypoxic conditions, respectively. Data represent the averages of three independent experiments (mean ± SD); *P < 0.05, **P < 0.01. EGR-1, early growth response factor-1; ELISA, enzyme-linked immunosorbent assay; H, hypoxia; N, normoxia.