Figure 3.

The effects on ASO activity were specific to depletion of La or NPM1. (A) Western analysis for La protein in cells treated with either siRNA or an ASO targeting La mRNA. γ-tubulin served as a loading control. UTC indicates mock-transfected cells. (B) Reduction of La levels by either siRNA or ASO caused similar effects on the activity of an ASO targeting PTEN mRNA. ASO treatment and qRT-PCR analysis were performed as described in Figure 2. (C) NPM1 protein was reduced by either an siRNA or an ASO targeting NPM1 mRNA as determined by western. γ-tubulin was used as a loading control. (D) Reduction of NPM1 protein by either siRNA or ASO decreased the activity of an ASO targeting PTEN mRNA. (E) La or NPM1 was transiently over-expressed in HeLa cells. After 48 h, protein levels were detected by western. (F, G) Over-expression of La or NPM1 protein increased the activity of ASOs targeting PTEN or NCL1 mRNAs, respectively. Where shown, the error bars are standard deviations of three independent experiments. P-values were calculated based on unpaired t-test (n = 3), and the significance is indicated above the bars. ‘NS’: not significant. ‘*’: 0.01<P<0.05; ‘**’: 0.001<P<0.01; ‘***’: 0.0001<P<0.001; and ‘****’: 0.00001<P<0.0001.