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. Author manuscript; available in PMC: 2014 Jul 23.
Published in final edited form as: Cell. 2010 Oct 1;143(1):46–58. doi: 10.1016/j.cell.2010.09.001

Figure 6. ncRNA-activators potentiate transcription of a reporter gene.

Figure 6

(A) ncRNA-a 3/4, 5 and 7 were cloned and inserted downstream of luciferase driven by a TK-promoter in a reporter plasmid as shown. (B) Graphical representation of the inserts in the various vectors used. The pGL3-TK-Control vector contains an insert of approximately 4 kb containing no annotated evidence of transcription. The depicted inserts show exons and transcriptional direction of the ncRNA-a. (C–E) Luciferase reporter assays. The Firefly luciferase vectors were co-transfected with a Renilla luciferase vector (pRL-TK) for transfection control. (C) The vector containing ncRNA-a3 and ncRNA-a4 from a bidirectional promoter, with control siRNA or siRNAs towards either of the two ncRNA-a, or both. (D) Reporter with ncRNA-5, and (E) the reporter with the ncRNA-a7 inserted downstream of luciferase. X-axes show relative Firefly (FL) to Renilla (RL) luciferase activity. Co-transfected siRNAs are indicated to the right of the bars. All data shown are from six independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 by one-tailed Student’s T-test.