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. 2009 Mar;15(3):459–472. doi: 10.1261/rna.1205409

FIGURE 3.

FIGURE 3.

Histone protein levels are decreased in SLBP knockdown cells, and the normal histone mRNA degradation in response to HU is impaired. (A) A serial dilution Western blot analysis of C2 (1:2–1:16, lanes 1–4) and a direct comparison of (lane 5) C2 and (lane 6) S2 total cell protein lysates. The levels of SLBP, histone H3 protein, and symplekin as a loading control were analyzed. (B) U2OS cells were knocked down for SLBP and nuclear proteins prepared from the control and SLBP knockdown cells. They were resolved on 15% SDS-polyacrylamide gels and stained with Coomassie blue. (Arrows) The core histone proteins. (C) Histone H2A mRNA is not degraded as efficiently in SLBP knockdown cells compared to control cells. HeLa cells were treated with (lanes 1–3) C2 or (lanes 4–6) S2 siRNAs and treated with 5 mM HU, and cells were harvested (lanes 2,5) 30 min or (lanes 3,6) 60 min later. (Left panel) Total cell RNA was analyzed by Northern blotting with probes to H2A mRNA and 7SK snRNA as a loading control. (Right panel) The results from three independent experiments. (Vertical bars) Standard deviations.