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. 2009 Apr 1;4(4):e5103. doi: 10.1371/journal.pone.0005103

Figure 4. Transcription from X mRNA Start site 2 can use either a free TBP or the TFIID complex.

Figure 4

(A) Examination of TBP concentration in HeLa NE. Indicated amounts of HeLa NE and purified recombinant TBP were loaded on a SDS-PAGE gel, and TBP was detected by anti-TBP western blotting. (B) In vitro transcription assays of the X gene (from Start site 2) and the Sp1-TATA templates (in a single two-template reaction). Control (lane 1) or TBP-depleted (lanes 2–8) NEs were tested for X gene or Sp1-TATA transcription in the absence (lane 2) or presence of 1 µl (lane 3) or 3 µl (lane 4) of purified TFIID or the presence of 1 ng (lane 5), 3 ng (lane 6), 10 ng (lane 7), or 30 ng (lane 8) of purified recombinant TBP (1 µl of the TFIID used in this experiment contained about 1 ng of TBP [6]). The enhancer-X promoter template was used to measure transcription from Start site 2. (C) Quantification results of the transcription assay. Transcription assays were performed twice for the reaction with 30 ng TBP (lane 8 of panel A) and four times for all of the other reactions. Brackets show standard errors of means.