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. 2012 Jan 13;7(1):e28916. doi: 10.1371/journal.pone.0028916

Figure 7. cDNAs of modified luciferase, hRluc and Luc2, do not mediate TPA-induced activation or negative regulation by T3/TR in CV1 cells.

Figure 7

A. Schematic representation of pBL-FFL-, pBL-hRluc-, and pBL-Luc2-CAT5. The cDNA of FFL, hRluc, or Luc2 was subcloned into the multi-cloning site of pBL-CAT5, which lacks pUC/AP-1 site. B. Co-transfection of Jun/Fos and/or treatment of TPA potentiated the enhancer activity of FFL cDNA, but not hRluc cDNA. pBL-FFL-CAT5 or pBL-hRluc-CAT5 (1.0 µg) was transfected into CV1 cells with or without the expression plasmids for Jun and Fos (0.2 µg each). After incubation for 24 h in the presence or absence of 100 ng/ml TPA, CAT activity was measured with normalization of the transfection efficiency by β-galactosidase activity. The results are the means +/− SD from three independent experiments. *, P<0.05. C. The cDNAs of hRluc [48] and Luc2 [51] do not function as TPA-induced enhancers and are not affected by T3/TR. TRβ1 expression vector (0.4 µg) was transfected into CV1 cells along with 1.0 µg of the pBL-FFL-, pBL-hRluc-, or pBL-Luc2-CAT5. After incubation for 24 h in the presence or absence of 100 ng/ml TPA and/or 1 µM T3, CAT activity was measured with normalization of the transfection efficiency by β-galactosidase activity. The results are the means +/− SD from three independent experiments. *, P<0.05.