FIG 9.

Tfeb and Tfe3 cis-activate a regulatory region of the Pparγ2 gene, and mutations of its E-boxes cause inhibition. (A) The murine Pparγ2 regulatory region (intron 1) construct. The fragment containing bp +132 to +4781 of the murine Pparγ2 intron 1 region was subcloned in the BamHI site of the pGl3 promoter vector. The positions of the E-boxes are +172, +203, +4792, and +4781. (B) 3T3-L1 preadipocytes were cotransfected with the SV40 promoter/luciferase vector containing the Pparγ2 regulatory region, the control Renilla, and the Tfeb and/or Tfe3 expression vectors. Luciferase activity was normalized to Renilla activity to control for transfection efficiency. Mutations to each E-box motif are shown (see Materials and Methods for sequences). The level of luciferase activity of the empty vector and in the absence of Tfeb and Tfe3 was defined by “1.” Fold activation was estimated to this level of activity. Values are means ± SD (n = 3).