Research Paper Volume 8, Issue 12 pp 3356—3374

Figure 2. Knockdown of NonO or PSF induces senescence. (A) Knockdown efficiencies for NonO siRNAs (si) and antisense (AS) cloned in pSuper-GFPNeo. Total RNA was isolated from 293T cells harvested 48 hr after calcium phosphate transfection with 10 μg either Si I or Si II silencing RNAs or with NonO AS DNA. Approximately 0.5 μg total RNA was reverse transcribed into cDNA which was analyzed by semi-quantitative PCR. (B) Growth reduction 72 hr post NonO knockdown as assessed by expression of pSuper-encoded GFP fluorescence microscopy at 1X and 10X magnification. (C) AS knockdown of NonO promotes senescence in Cos7 cells. G418-selected Neo^R clones of Cos7 or HeLa transfectants assayed at the indicated population doubling (PD) by SA-β-gal staining. (D) Overexpression (OE) or knockdown (AS) of PSF expression induces senescence in Cos7 or HeLa cells analyzed in bulk 96 hr following transfection.