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. 2011 May 2;108(20):8233–8238. doi: 10.1073/pnas.1017700108

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Fig. 5. — Dephosphorylated RS domain and U1-70K binding defective mutant of SRSF1 block early spliceosomal assembly steps. (A) Cartoon showing β-gb (Ron) and β-gb (Ron)ΔAG constructs. (B) Native gel analysis of the spliceosomal E complex formation by WT SRSF1 (RSRS) and different SRSF1 phosphomimetics in the presence of S100 extract and β-gb (Ron) pre-mRNA substrate. (C) Native gel analysis of the spliceosomal A and B/C complex formation of β-gb (Ron)ΔAG by SRSF1 phosphomimetics and WT (RSRS) without (left) or with 0.1 mg/mL tRNA (right) (D) In vitro splicing of the β-gb (Ron) pre-mRNA substrate by SRSF1 phosphomimetics in the presence of S100 extract. The relative splicing activities are shown at the bottom. (E) Native gel analysis of the E complex formation by WT and mutant SRSF1 (RRM1/2) in S100 extract. (F) Native gel analysis of the spliceosomal A and B/C complex formation of β-gb (Ron)ΔAG by WT and SRSF1 (RRM1/2) mutants without (left) or with 0.1 mg/mL tRNA (right) in S100 extract. (G) In vitro splicing assay of β-gb (Ron) pre-mRNA by WT and mutants SRSF1 (RRM1/2) in S100 extract. The relative splicing activities of SRSF1 (RRM1/2) mutants compared to WT as shown at bottom.