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. 2003 Nov 1;31(21):6104–6116. doi: 10.1093/nar/gkg817

Figure 5.

Figure 5

Splicing inhibition by peptides derived from the PLRG1 binding region of CDC5L. (A) Double-headed arrows indicate peptides synthesised from the PLRG1 binding region of CDC5L. (B) Autoradiograph of a splicing gel using CDC5L 24mer peptides. The peptides were added to splicing reactions to check the effect on splicing. Lane 1, the input pre-mRNA; lane 2 (CTRL1), a splicing reaction without peptides; lane 3 (CTRL2), a negative control reaction containing the peptide CD-R24 (CD-R24 contains the same amino acids as the peptide CD24-1 but in a different order); lanes 4–6, peptides CD24-1, CD24-2 and CD24-3, respectively. (C) Effect of the CDC5L inhibitory peptide on splicing using other pre-mRNA templates (AL4, hnRNP A1 and RPL32). Lanes 1, 6 and 11, the input pre-mRNA; lanes marked CTRL1, splicing controls without peptide; lanes marked CTRL2, control peptide CD-R24. The lanes with splicing reactions containing CD24-2 (i.e. lanes 5, 10 and 15) also served as internal controls because the peptide is also derived from the PLRG1 binding region of CDC5L. Lanes 4, 9 and 14 are splicing reactions that contained the CD24-1 peptide. Note that the size differences between the introns of these constructs have not been resolved by this gel because it was not run for the amount of time needed to resolve these size differences. (D) Effect of 8mer–12mer peptides derived from the CD24-1 sequence on splicing. Lane 1, input pre-mRNA; lane 2 (CTRL1), a splicing reaction without peptide; lane 3 (CTRL2), a splicing reaction containing the control peptide CD-R12 (CD-R12 contains the same amino acids as CD12-3 except that the order of the amino acids in the sequence is different); lanes 4–6, splicing reactions that included the 12mer peptides CD12-1, CD12-2 and CD12-3, respectively. The splicing reaction in lane 7 contained the 8mer peptide CD8. The symbols on the right of the panel represent the input RNA, splicing intermediates and products. The other bands in the figure have been caused by partial degradation of the RNA in the reactions.