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. 2018 Feb 5;46(7):3774–3790. doi: 10.1093/nar/gky070

Figure 4.

Figure 4.

Transiently-expressed, truncated snRNAs containing Sm and SMN binding sites accumulate in Cajal bodies. (A) Immunoprecipitation of U2-MS2 and the deletion mutant U2ΔSLI+IIa,b-MS2. RNAs were immunoprecipitated via MS2-YFP by anti-GFP antibodies and detected by silver staining. Co-precipitated proteins were analyzed by Western blotting. With the full-length U2-MS2 RNA we detected all tested proteins (SmB/B′, SF3A3, SNRPB2 and SF3B4), while only SmB/B′ and SNRPB2 co-precipitated with the U2ΔSLI+IIa,b-MS2 RNA. (B) Immunoprecipitation of U4-MS2 and the deletion mutant U4Δ1–64-MS2. Legend as in (A). With the full-length U4-MS2 RNA we detected both tested proteins (SmB/B′ and PRPF31), while only SmB/B′ co-precipitated with U4Δ1–64-MS2 RNA. (C) Hela cells were co-transfected with U2 or U4 snRNAs containing the MS2 loop (green stem loop) and MS2-YFP (green). Coilin was used as a marker of CBs (red). Small red box in snRNA schemes represents the canonical Sm site and blue boxes represent endogenous U2 or U4 promoters. DNA was stained by DAPI. The scale bar represents 10 μm.