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. 2004 Mar;78(5):2201–2211. doi: 10.1128/JVI.78.5.2201-2211.2004

FIG. 2.

FIG. 2.

The GABP-binding sites are required, together with the GRE, for the TGF-β superinduction of the MMTV promoter (A, B, and C) and of the SV40 promoter (D and E). Transient transfection and treatment of M12 cells were as described in the legend to Fig. 1B. (A to C) pGL3-basic containing the following LTRs: wild type (wt LTR or Lwt); LTR mutated in fp1/fp2 (LTR mut. fp1/fp2 [see Fig. 1A]); LTR 5′ truncated at −303, wild type (-303 wt), or LTR mutated in fp1/fp2 (-303 mut. fp1/fp2); LTR mutated only in fp1 (Lmut fp1) or in fp2 (Lmut fp2); a promoter-negative LTR deleted from positions −105 to +133 (ΔP1), as a negative control. (D and E) pGL3-(SV40)-promoter containing the indicated number of head-to-tail boxes (nb. box) with the wild-type sequence from positions −116 to −209 (2i = two in inverted orientation) (D) or none (−), or else two boxes, either wild type (wt) or mutated in the GRE (mutGRE = LS−175/−166 [10]), in fp1/fp2 (mut fp1,2), or in both GRE and fp1/fp2 (mutGRE + fp1,2) (E). The negative control was as in panel C. Cotransfected internal standards (50 ng) were pRL-SV40 in panels A and B and pRL-SV40Δ enhancer in panels C, D, and E. Error bars indicate the standard deviations. Experiments were repeated at least twice.