FIGURE 7.

TRIP12 reverses gene activation and antiproliferative activity of PTF1a. A, HEK-293T cells were transiently co-transfected with PTF1 complex (composed of PTF1a, RBPL, and HEB), the 6XA26-luc reporter construct that contains an hexamer of PTF1-responsive promoter and RSLV40 vector, in the presence of increasing amounts of TRIP12 (100, 200, and 500 ng/well) (black bars), TRIP12-C1959A (200 and 500 ng/well) (gray bars) or control pDEST47 plasmid (200 and 500 ng/well) (white bars). The RSLV40 vector was included as an internal control for normalization. The results are expressed as percentages of activation (means ± S.E., n = 3), and each experiment is performed in triplicate. No significant differences on the transcriptional activity are observed when TRIP12 or TRIP-C1959A constructs were transfected in the absence of PTF1a (data not shown). B, HEK-293T cells were co-transfected with PTF1a and with TRIP12 or TRIP12-C1959A or their corresponding controls plasmids (as indicated), stopped, and counted 6 days post-transfection. The results of experiments performed in triplicate are represented as counted cells per well and expressed in means ± S.E. (n = 3).