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. 1997 Jun 10;94(12):6324–6329. doi: 10.1073/pnas.94.12.6324

Figure 4.

Figure 4

Altered specificity TBP assay in HeLa cells to test the effect of TBP mutations H1′+H2+S3′/S4′ and H1′+S3/S4+S3′/S4′ on the transactivation potential of CIITA activation domain. Histograms represent the following transfections. Lane 1; transfections to check the levels of CIITA-mediated activation due to endogenous TBP. c-fos(-56)TGTAAAA(4xgal) reporter construct (2 μg) and 320 ng pf PBGX1CIITA(1–210) effector construct was used for each transfection. The same amounts of the reporter and effector plasmids were used for the remaining transfections. Lane 2; transfections to check the levels of CIITA-mediated activation achieved by the addition of 224 ng of pCGNhTBPAS-WT, the expression plasmid for the altered specificity TBP. Also shown are transfections with the mutant altered specificity TBP molecules H1′+H2+S3′/S4′ (560 ng, lane 3) and H1′+S3/S4+S3′/S4′ (560 ng, lane 4), respectively. These amounts of the two combinatorial TBP mutants were previously determined (36) by quantitative Western blotting to result in expression levels equivalent to that obtained with 224 ng of TBPAS-WT plasmid.