Figure 6
PPARγ binds to PPRE3 in human MnSOD promoter. (A) Schematic representations of full length human MnSOD promoter construct (-3400 to +24 pGL3) and two 5'-deletion constructs (-1605 to +24 pGL3; -555 to +24 pGL3) with location of PPRE1, PPRE2 and PPRE3 indicated. (B) Luciferase assay of MnSOD promoter activity in MDA-MB-231 transiently transfected with full length and 5'-deletion derivatives followed 5 μM and 10 μM 15 d-PGJ2 treatment for 16 h. Renilla plasmid was cotransfected to assess transfection efficiency. Results are expressed as RLU/renilla/μg total protein. Data represents the average +/-SD of two experiments done in duplicate. (C) No-shift DNA binding assay to assess DNA binding activity of PPARγ at PPRE2 and PPRE3 of MnSOD promoter as described in M&M. Results are expressed as absorbance readings at 450 nm. Data represents the average +/- SD of two experiments done in duplicate.