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. 2006 Oct 30;103(45):16977–16982. doi: 10.1073/pnas.0601565103

Fig. 4.

Fig. 4.

Dach2 inhibits Mgn promoter activity and Dach2 knockdown abrogates TSA-mediated Mgn promoter inhibition. (A) Dach2 overexpression inhibits Mgn promoter activity. C2C12 cells were cotransfected with the 133-bp Mgn promoter-luciferase expression vector, CMV-CAT for normalization, and either pCS2GFP or pCDNA3Dach2. Cells were differentiated, and 4 days posttransfection, myotubes were harvested for luciferase and CAT assays (n = 3). (B) Dach2 knockdown rescues Mgn gene expression in myotubes treated with TSA. C2C12 cultures were transfected with the 133-bp Mgn promoter driving luciferase expression along with CMV-CAT, and with either a control or two different Dach2-targeted siRNAs (1# and 2#). Transfected and differentiated myotubes were treated with either TSA or vehicle for 12 h before harvesting for luciferase and CAT assays (n = 3). (C and D) The MEF3 element of the Mgn promoter confers Dach2-dependent suppression on the minimal enkephalin promoter (MEK). HEK cells (C) (n = 3) or adult denervated muscle (D) (n = 4) were transfected or electroporated, respectively, with the indicated vectors. Three days later, cells or tissue were harvested for luciferase and CAT assays. Error bars are standard deviation.