Figure 2.

HuB and HuD associate with TERC in the same motif. (A) Schematic representation depicts the fragments of TERC used for RNA pulldown assays. (B) RNA pulldown assays were performed using SH-SY5Y cell lysates and in vitro-transcribed TERC fragments depicted in (A). The CR (coding region) and 3′UTR fragments of p27 mRNA served as negative (N) and positive (P) controls, respectively. A 5-μg aliquot input (Inp.) and binding to GAPDH were also assessed. (C) Schematic representation depicts full length TERC (WT) and its variants used for pulldown assays. (D) The association of HuB and HuD with TERC variants bearing mutations U40A, U100A, or U40A + U100A (U40/100A) (Figure 2C, Schematic) was determined by using RNA pulldown assays. (E) Quantification of the bands detected by western blot analysis in (D); data are the means ± SD of the signals from three independent experiments and significance is analyzed by two-tailed Student's t test (**P < 0.01).