Fig. 1.

Coilin interaction with the telomerase complex protein dyskerin is mediated by hTR. (A) Lysate was immunoprecipitated (IPed) with either control (rabbit IgG) or coilin polyclonal antibodies, followed by running the samples on SDS-PAGE and western transfer. The probing antibodies are indicated. (B) Lysate was IPed with control (mouse IgG), dyskerin monoclonal or dyskerin polyclonal antibodies. Probing antibodies are indicated (Hc in lane 4=IgG heavy chain). (C) Lysate was IPed with control (rabbit IgG) or coilin polyclonal antibodies. Antibody-bead complexes were untreated (−) or treated (+) with RNase. Probing antibodies are indicated. (D) Cells were transfected with two different hTR siRNAs, followed by IP with anti-coilin antibodies. Probing antibodies are indicated. Quantification of dykserin and SMN recovery relative to that obtained with control siRNA for this and other experimental replicates is shown in the lower panel. For all IPs, inputs represent 1.5% of the cell lysate used in the IP reactions. (E) HeLa cells were treated with control or hTR siRNA (hTR #1 or hTR #2) for 48 h. Total RNA was collected and the amount of hTR was analyzed by qRT-PCR, relative to GAPDH message. The level of hTR in cells treated with hTR siRNA was compared to that found in cells with control siRNA for each time point (n=3). *P<0.05 compared to control knockdown. For all histograms, error bars denote standard error about the mean.