Figure 5.

H₂O₂ induces TR4 gene expression through activating FOXO3a. A, C2C12 cells (1 × 10⁴) were plated into two-well chamber slides for 24 h and then untreated or treated with 250 μm H₂O₂ for 2 h. Cells were fixed and stained as described in Materials and Methods. Con, Control; DAPI, 4′,6′-diamidino-2-phenylindole. B, pGL-DBE × 6 (500 ng) were transfected into H1299 and C2C12 cells for 40 h and then left untreated or treated with 250 μm H₂O₂ for 2 h; the Luc activity was measured 6 h after H₂O₂ treatment. **, P < 0.01 (Student’s t test analysis) as compared with control. C, pGL-TR4-1920 (500 ng) were transfected into H1299 and C2C12 cells for 40 h and then left untreated or treated with 250 μm H₂O₂ for 2 h; the Luc activity was measured 6 h after H₂O₂ treatment. *, P < 0.05 (Student’s t test analysis) as compared with control. D, Total cell lysates of C2C12, H1299, and primary MDF cells untreated or treated with 250 μm H₂O₂ for 2 h were analyzed for expression of TR4 with actin as a loading control. E, Total RNA isolated from C2C12, H1299 and primary MDF cells untreated or treated with 250 μm H₂O₂ for 2 h were subjected to real-time PCR. **, P < 0.01 (Student’s t test analysis) as compared with control.