Fig. 5.

Hypoxia-related miRNA expression. (A) Differentially expressed miRNAs in hypoxic 3T3-L1 adipocytes vs normoxic cells. miRNAs whose expression was either upregulated (> 2-fold) or downregulated (< 0.5) after hypoxia treatment (1% O₂) of cells are shown in the diagram (left). The expression levels of miR-128 in normoxic and hypoxic 3T3-L1 adipocytes were determined by RT-qPCR (right); results are the means ± s.e.m. from 3 independent experiments, each in triplicate; P < 0.001 vs normoxia [Student's t-test]. (B) miR-128 expression in VAT from normal-weight and obese individuals (n = 10 per each group), and mice under NCD- and HFD-fed conditions (n = 10 per each group) for 15 weeks. In the obese subject category, VAT samples were as follows: 3 (BMI 30–34.9); 3 (BMI 35–39.9); 4 (BMI ≥ 40). Data are the means ± s.e.m. of 2 independent RT-qPCR assays from each individual tissue sample. *P < 0.05 vs control (white bar) [Student's t-test]. (C) miR-128 levels in liver and skeletal muscle from NCD- and HFD-fed mice as measured in (B). (D) miR-128 levels as measured by RT-qPCR in VAT from normal-weight, non-obese individuals (n = 10), placed in organ culture for 48 h, either in normoxic or hypoxic environment, or after reoxygenation. P < 0.001 vs normoxia [Student's t-test]. (E) Normoxic 3T3-L1 adipocytes were transiently transfected with an effector plasmid (1 µg) expressing HIF-1α. After 48 h, miR-128 levels were measured by RT-qPCR. Results are the mean ± s.e.m. of triplicates from 3 independent assays. A representative WB of HIF-1α is shown for each condition. White bar, mock (no DNA); black bar, pcDNA3-vector without an insert; gray bar, pcDNA3-HIF-1α effector vector.