Fig 4.
Analysis of transcription factors assembled with ERV-9 LTR U3 RNAs by RNA-IP. A total of 5 × 107 cells from tumor cell lines and stimulated or unstimulated primary cells were used for RNA-IP (RIP) analysis. The cell lysates were treated with antibodies to each of the following: NF-Y, p53, p300, Ets-1, Sp1, HDAC1, IgG (nonspecific negative control shown in Fig. S2 in the supplemental material), and p21/WAF1 (transcription factor-specific negative control), as well as antibody to PTBP1 as a loading control. The RT-PCR detection of ERV-9 LTR U3 region is described in the legend to Fig. 1b. NF-Y, p53, and sp1 assemble with U3 AS RNAs in HT1080 and MDA231 cells and in quiescent and stimulated human primary T cells.