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. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Cancer Res. 2009 Dec 22;70(1):229–239. doi: 10.1158/0008-5472.CAN-09-2788

Figure 4.

Figure 4

Sam68 binds to intron 4 sequences in the cyclin D1 pre-mRNA. A, scheme of the CCND1 gene comprised between exon 4 (E4) and exon 5 (E5). White box, position of the Sam68-binding sites in intron 4 (int4). Arrows, genomic regions amplified in the ChIP experiment. Cross-linked chromatin derived from LNCaP cells was immunoprecipitated and analyzed by quantitative real-time PCR using the indicated primers as described in Fig. 1. Columns, mean of three independent experiments; bars, SD. P values of Student's t test are reported. B, pull-down assays of Sam68, U1-70K, and hnRNP A1 with biotinylated RNA containing CCND1 exon 4 and different regions of intron 4 with (Rev1) or without (Rev2 and Rev3) the potential Sam68-binding sites. The proteins pulled down with the RNAs were analyzed by Western blot with antibodies for Sam68, U1-70K, or hnRNP A1.