Methylated TLS binds CCND1-pncRNA-D. RIP lysate from HeLa cells were immunoprecipitated using either 2B12 or a normal mouse IgG as a negative control. RNA associated with methylated TLS was purified, and validated by RT-PCR using the specific primers for CCND1-pncRNA-D. The PCR prodcuts were ran on an agarose gel to detect the presence of CCND1-pncRNA-D. The “input” omits the immunoprecipitation step, “IgG” used an IgG antibody for the immunoprecipitation, “2B12” used a 2B12 antibody to pull down methylated TLS, and “water” lane served as a negative control for the PCR reaction. The expected size of PCR product for CCND1-pncRNA-D could be detected in 2B12 RIP. PCR product was observed in the 10% input and not in the normal mouse IgG RIP.