P-FMRP is unable to associate with Dicer. (A) Lysate from 2 × 107 HeLa cells was immunoprecipitated with NP or PSER antibodies and immunoblotted (ib) with Dicer antibody or FMRP (1a) antibody. (WCL)-50 μg HeLa whole-cell lysate; IP, immunoprecipitations; and Ig, immunoprecipitating antibody alone. (B) Lysate from 109 Flag-FMRP-expressing cells was immunoprecipitated with anti-FMRP antibody 7G1 (Brown et al. 2001), split, and incubated with either RNase (+) or mock-treated (−) and immunoblotted (ib) with the Dicer antibody. (WCL)-50 μg whole-cell lysate. (C) Lysate from 107 HeLa cells was incubated with matrix-coupled peptide (NH2-SNA[pS]ETES-CONH2) (P-FMRP), (NH2-SNASETES-CONH2) (FMRP), or random peptide (RP) (KETAAAKFERQHMDS); resolved on a 6% SDS-PAGE gel; and immunoblotted with Dicer antibody.
