Fig. S2
Differential Activity of the Minor and the Major Spliceosome on an In Vitro-Synthesized Pre-mRNA Localized in the Cytoplasm
In vitro transcribed, capped and polyadenylated human (Hu)P120 minigene pre-mRNA, schematically shown in panel (A), was electroporated into asynchronously growing mouse NIH 3T3 cells. (B) RT-PCR analysis of splicing of minor-class intron F and major-class intron G from the transfected transcript (6h after transfection); for primer positions, see panel (A). -RT, corresponds to RT-PCR reactions lacking reverse transcriptase. Correct splicing was verified by sequencing. (C) Detection of the transfected pre-mRNA by fluorescence in situ hybridization in transfected (upper panel) and mock-transfected cells (lower panel). A DIG-labeled LNA oligonucleotide specific for to human P120 intron G was used as a probe. Nuclear/DNA counterstaining was performed with DRAQ 5.
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