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. 2019 Mar 13;47(9):4778–4797. doi: 10.1093/nar/gkz168

Figure 13.

Figure 13.

Tubercidin causes an mRNA export defect. (A) RNA FISH experiment to detect the distribution of the E6 mRNA in U2OS cells treated with tubercidin and Pladienolide B (PlaB) using a Cy5 labeled probe that detects the MS2 region of the E6 mRNA (yellow), and a Cy3-labeled probe that detects the intron of the E6 mini-gene (red). SGs were detected using an anti-G3BP1 antibody (magenta). Hoechst DNA stain is in blue. (B) Splicing inhibition causes RNA retention in nuclear speckles. The localization of E6 transcripts under splicing inhibition by Pladienolide B treatment in U2OS cell was followed by RNA FISH using a Cy5-labeled MS2 probe (green). Nuclear speckles were detected using anti-SRSF2 (red). Hoechst DNA stain is in blue. Bar = 5 μm. (C) Semi-quantitative RT-PCR analysis of cells treated with Pladienolide B (10 μM) or tubercidin (10 μM) together with dox induction for 6 h. Cells were examined for intron inclusion of HSP40 and DXO pre-mRNAs and for exon skipping of E6, MCL1, NOP56 and p27 pre-mRNAs. The positions of different cDNA products are noted on the right of the gel images.