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. 2017 Jun 5;216(6):1579–1596. doi: 10.1083/jcb.201701165

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© 2017 Malinová et al.

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Figure 3. — RP mutations inhibit U5 snRNP assembly. (A) Association of PRPF8 variants with U5 and U4/U6.U5 snRNPs. PRPF8-LAP variants are indicated at the top. IP was done with anti-GFP antibodies. Inputs contained 2% of the materials in the IP, and the antibodies used for WB are indicated at the right. (B) Quantification of the immunoprecipitated material. The graph displays the amount of the protein present in the pellet and normalized, first to the amount of copelleted GFP and then to the same ratio measured in the PRPF8-WT-LAP pellets. (C and D) Association of PRPF8 variants with AAR2. Legend is as in A and B. Error bars represent SEM. The exact number of each experiment is indicated below/in each column. *, P < 0.05, as measured with t tests comparing the indicated sample and WT PRPF8-LAP IPs. (E) Association of three selected PRPF8 variants with snRNPs. WBs were made with extracts of cells expressing the PRPF8-LAP variant indicated at the top (input lanes) and immunoprecipitated with anti-Sm Y12 antibody. PRPF8-LAP proteins were detected with anti-GFP antibody and SNRPB/B’ with the Y12 antibody. Inputs contain 2% of the materials in the IP.