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. 2006 Jul 28;7(9):898–903. doi: 10.1038/sj.embor.7400767

Figure 2.

Figure 2

Distinct effect of transcriptionally defective TFIIB mutants in vivo. (A) Plasmids driving the expression of wild-type TFIIB (wtIIB) or the indicated mutants (2 μg) were introduced into embryonic kidney 293 cells with 2 μg of the reporter G9E4CAT (top) or HIV LTR CAT (bottom) and a plasmid driving the expression of GAL4(1–94)-VP16 (0.2 μg). After 48 h, RNA was prepared and CAT transcripts were detected by primer extension and autoradiography. (B) A 293-derivative cell line that contains G9ML-Luciferase stably integrated was transfected as in (A), but without the reporter. After 48 h, cells were lysed and luciferase activity was measured and is presented graphically. A blot (anti-T7) showing expression of the TFIIB derivatives is shown at top right. CAT, chloramphenicol acetyltransferase; HIV LTR, human immunodeficiency virus long terminal repeat; TFIIB, general transcription factor IIB.