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. Author manuscript; available in PMC: 2010 Oct 25.
Published in final edited form as: Nat Methods. 2009 Oct 4;6(11):825–830. doi: 10.1038/nmeth.1379

Fig. 3. Various RS domains or Gly-rich domains can function as the effector module of ESFs.

Fig. 3

(a) Exon-skipping reporters containing the 8-nt cognate sequence of PUF3-2 ESFs in the cassette exon were used to examine the splicing suppressor activities of Gly-rich domains/fragment (left panel) or the splicing activator activities of RS domains/fragment (right panel). The experiments were similar to Fig. 1d and 1g except that different functional domains were fused to the same PUF3-2. (b) Inhibition (lanes 1-4) or promotion (lanes 6-10) of exon inclusion by ESFs with different splicing effector domains. The experiment conditions were same as Fig. 1d (for Gly-rich domains/fragment) or Fig. 1g (for RS domains/fragment), and all transfections were repeated at least twice. The functional domains of ESFs are lane 1 to 3, the Gly-rich domains of hnRNP A1 family proteins; lane 4, a short peptide of Gly-rich sequence (GYGGGGPGYGNQGGGYGGG); lane 6-9, the RS domains of SR proteins; lane 10, a short RS repeat (RS)6. (c) Fold change of cassette exon inclusion relative to the reporter alone (lane 5, no ESF). The means of replicated experiments are plotted with error bars indicating the range of replicated experiments.