ZFIN Image: König et al., 2007, Fig. 3

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Figure Caption

Fig. 3 Minor Splicing Evades Mitotic Downregulation

(A) Scheme of the P120 pre-mRNA region analyzed for splicing (boxes, exons; lines, introns; arrows, primers).

(B) Flow cytometry analysis of cell DNA content from NIH 3T3 cells that grew asynchronously (cycling cells) or were arrested in mitosis by nocodazole treatment.

(C) RT-PCR analysis of U2-type (introns E and G) and U12-type (intron F) splicing of transfected, in-vitro-transcribed P120 minigene pre-mRNA (capped and polyadenylated) in mitotic cells, after the times indicated. For primer positions, see (A). -RT corresponds to RT-PCR reaction lacking reverse transcriptase. Correct splicing was verified by sequencing.

(D) Analysis of splicing of endogenous P120 pre-mRNA in cycling (label C) and mitotically arrested cells (label M). Relative expression levels of mRNA spliced for the major class introns (In) G and E and for the minor-class intron F were measured by real-time quantitative RT-PCR. Values represent the mean of two experiments and are expressed as relative percentage of unspliced RNA (set to 100% in cycling cells).

Acknowledgments

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Reprinted from Cell, 131(4), König, H., Matter, N., Bader, R., Thiele, W., and Müller, F., Splicing Segregation: The Minor Spliceosome Acts outside the Nucleus and Controls Cell Proliferation, 718-729, Copyright (2007) with permission from Elsevier. Full text @ Cell