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. 2013 Oct 31;42(3):1893–1903. doi: 10.1093/nar/gkt1020

Figure 5.

Figure 5.

Access to exon 11 sequences is not required for MBNL1-mediated activation of upstream intron splicing or U2AF65 binding. (A) The upstream intron RNA substrate was spliced in the presence of an exon 11-spanning (E11block) or control (ctrl) MO and visualized by autoradiography. The location of the exon 11 MO is indicated on the right. The ∼240-nt band appears in some experiments using both MOs in the presence and absence of ATP, suggesting they are non-specific degradation products (data not shown). (B) Quantification of the MBNL1-induced fold change in lariat intermediate formation using the phosphorimager signal for the percentage lariat intermediate (compared with total RNA isoforms in the lane). (*P < 0.05) (C) UV cross-linking followed by immunoprecipitation for U2AF65 (as described in Figure 4B) in the presence of the exon 11 MO (E11). An autoradiogram (AR) and western blot for U2AF65 (WB) are shown. The bottom panel shows quantification of the MBNL1-induced fold change in RNA binding (normalized to protein levels as in 4B, *P < 0.05).