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. Author manuscript; available in PMC: 2009 Mar 15.
Published in final edited form as: Cancer Res. 2008 Sep 15;68(18):7621–7628. doi: 10.1158/0008-5472.CAN-08-1217

Figure 4. SNARP complex proteins are upregulated in G1 phase and are required for Cyclin D1 mRNA production.

Figure 4

(A) Cell cycle analysis of SNIP1 associated proteins. U-2 OS cells were fractionated by centrifugal elutriation, whole cell extracts prepared and equivalent protein amounts separated by SDS-PAGE and subjected to western blot analysis using the indicated antibodies. Cells from each fraction were also stained with propidium iodide for analysis by FACS and cell cycle percentages of each fraction are shown graphically.

(B) Analysis of RNA transcript levels following depletion of SNARP complex components. U-2 OS cells were treated with siRNAs directed against SNIP1, SkIP and THRAP3/BCLAF1 (knocked down together due to their sequence similarity and possible functional redundancy). The relative levels of pre-spliced and post-spliced RNAs were then assessed by qRT-PCR, normalised to GAPDH levels. The mRNA (post-spliced) : hnRNA (pre-spliced) ratio for the Cyclin D1 transcript is also presented. The pre-spliced transcript is detected using primers within intron 1 and the post-spliced transcript detected using primers situated within exons 3 and 5.