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. 2008 Jul 16;3(7):e2721. doi: 10.1371/journal.pone.0002721

Figure 2. Specific induction of TAF6δ protein expression by modified antisense RNA oligonucleotides.

Figure 2

(A) HeLa cells were transfected with splice site-switching oligonucleotides directed against exon IIα of the TAF6 gene and treated with MG-132 5 hours later. 21 hours post transfection cells were fixed and stained with the indicated antibodies for immunocytochemistry (B) Quantification of endogenous TAF6δ expressing cells transfected with splice site-switching antisense oligonucleotides. Results are expressed as the percentage of cells displaying a clear TAF6δ punctate staining on a total of at least 500 cells. (C) Translation of exogenous TAF6δ is induced by modified antisense RNA oligonucleotides. Schematic representation of plasmid pASTAF6 containing sequences derived from the TAF6 cDNA (white) or from genomic DNA (grey). HeLa cells were first transfected with pASTAF6 and 19 hours later with splice site switching oligonucleotides and treated with MG132 and ZVAD-FMK 6 hours after this second transfection. 38 hours post-transfection protein extracts from cells were analyzed by immunoblotting with monoclonal antibodies directed against TFIID subunits.