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. 2008 Oct;19(10):4141–4153. doi: 10.1091/mbc.E07-12-1217

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© 2008 by The American Society for Cell Biology

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Figure 3. — Down-regulation of YY1 expression results in up-regulation of the fetal isoforms and in increase in cell size and protein synthesis. (A) YY1 expression is repressed by transfection with an YY1 siRNA oligonucleotide (lane 3) but not by a control or an unrelated siRNA oligonucleotide (lanes 2 and 4). Lane 1, no transfection. Total extract was used, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. (B) Cells were transfected with YY1 siRNA nucleotide and treated with vehicle (white bars) or PE (black bars) for 48 h. RNA expression was analyzed by RT-PCR and normalized to 18S. Results were compared with vehicle-treated cells transfected with a control siRNA, defined as 100%. n = 4. (C) Radiolabeled protein experiments show an increase in radiolabeled leucine incorporation in cells transfected with YY1 siRNA oligonucleotide. (D) Cell size measurements show an increase in cell size in cells that do not express YY1. Fifty-five cells from three different experiments were measured. (E) Immunofluorescence with anti-actinin antibody of cells transfected with a control or YY1 siRNA oligonucleotide.

Figure 3. — Down-regulation of YY1 expression results in up-regulation of the fetal isoforms and in increase in cell size and protein synthesis. (A) YY1 expression is repressed by transfection with an YY1 siRNA oligonucleotide (lane 3) but not by a control or an unrelated siRNA oligonucleotide (lanes 2 and 4). Lane 1, no transfection. Total extract was used, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. (B) Cells were transfected with YY1 siRNA nucleotide and treated with vehicle (white bars) or PE (black bars) for 48 h. RNA expression was analyzed by RT-PCR and normalized to 18S. Results were compared with vehicle-treated cells transfected with a control siRNA, defined as 100%. n = 4. (C) Radiolabeled protein experiments show an increase in radiolabeled leucine incorporation in cells transfected with YY1 siRNA oligonucleotide. (D) Cell size measurements show an increase in cell size in cells that do not express YY1. Fifty-five cells from three different experiments were measured. (E) Immunofluorescence with anti-actinin antibody of cells transfected with a control or YY1 siRNA oligonucleotide.