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. 2001 Feb 1;29(3):732–742. doi: 10.1093/nar/29.3.732

Figure 1.

Figure 1

TGF-β responsive co-modulators, Smad2 and 4, increase the activities of both GAL4–MEF2A and GAL4–MEF2C. (A and B) C2C12 cells were co-transfected with pCMV5B–TβRI(T204D), pCMV5B–Smad2, pCMV5B–Smad4, 5×GAL4-luc, GAL4(DBD), GAL4–MEF2A, GAL4–MEF2C and pSV-βgal as indicated and treated with (+) or without (–) 2 ng/ml of TGF-β in 1% FBS. The transcriptional activation domains of both MEF2 isoforms are fused to the DNA binding domain (DBD) of GAL4 to form GAL4–MEF2A (91–507 amino acids) and GAL4–MEF2C (87–442 amino acids). GAL4(DBD) contains the GAL4 DBD only. The reporter plasmid, 5×GAL4-luc, contains five copies of the GAL4 binding site linked to the adenovirus EIB promoter and the firefly luciferase gene. After 72 h, β-galactosidase activities were measured and used to normalise luciferase activity values. Two or more sets of assays were performed in both COS and C2C12 cells with comparable results. Each data point is a mean of triplicate samples from single experiments and the error bars represent the standard error of the mean (SEM).