Figure 3.
U11-59K (PDCD7) mediates exon-definition interactions between the major and minor spliceosomes. (A) Bar graph showing the %MSIAS of the Mlst8 splicing reporter, in response to siRNA against splicing factors. Inset shows schematic of the Mlst8 splicing reporter and the resulting products. Agarose gel image of the Mlst8 splicing reporter RT-PCR products resulting from downregulation of minor spliceosome proteins (B) and quantification of the %MSIAS using ImageJ (C). Significance was determined using one-way ANOVA, followed by the post-hoc Tukey test. (D) Agarose gel images of RT-PCR products resulting from AS around minor introns upon knockdown of PDCD7 in A549 cells. (E) Heatmap of the exclusive unique spectrum count of the Pdcd7-interacting proteins involved in splicing. (F) Immunoblot for major spliceosome proteins that immunoprecipitated with Pdcd7. (G) Immunoblot for PDCD7 after immunoprecipitation of endogenous U2AF1. (H) Immunoblot for major spliceosome proteins that immunoprecipitated with Pdcd7 with and without RNase A treatment. Arrow points at predicted molecular weight of U2AF1. Asterisk marks predicted molecular weight of U2AF2. *P < 0.05; **P < 0.01. See also Supplementary Figures S4–S6.