Figure 3

The effect of Sqle on adipogenesis. (A) 3T3-L1 cells were seeded on 24-well plate. After 24 h, cells were transfected with control and Sqle siRNA. When the confluency of cells reached 70%, at which point 3T3-L1 cells were differentiated in adipogenic differentiation media containing DMEM supplemented with 10 µg/mL insulin, 0.5 mM 3-isobutyl-1-methylxanthine (IBM-X), and 1 μM dexamethasone. Cells were fully differentiated into adipocytes after 7 d, at which point cells were fixed in 10% (v/v) formaldehyde in PBS and stained with Oil Red O solution. Stained cells were observed under a microscope. (B–D) Cells were seeded on 6-well plate. After 24 h, cells were transfected with control and Sqle siRNA. The adipogenic differentiation was induced as described above. The RNA from undifferentiated cells and differentiated cells transfected with control and Sqle siRNA was prepared and subjected to RT-qPCR. Gene expression levels (B;Pparg, C;Cebpa, D;Adipoq, E;Lep) were analysed using the 2^−∆∆Cq method. Gapdh was used as an internal control gene. Fold change was calculated by dividing expression levels in the experimental group by those of the undifferentiated group. All experiments were done at least three repeats. The significant differences were shown as *P < 0.05 versus undifferentiated cells and ^# P < 0.05 versus control siRNA transfection.