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. 2012 Oct 1;109(42):16841–16846. doi: 10.1073/pnas.1215406109

Fig. 3.

Fig. 3.

WNK3 regulates splicing activity of Fox-1 and Fox-2. (A) Genomic organization and alternative splicing patterns of an endogenous FMNL3 region including exons 25, 25a, and 26 and introns (Upper). FMNL3 minigene content (Lower). The region marked with dotted lines was inserted between β-globin exons 1 and 2 in the pcAT7-Glo1 plasmid. (B) FMNL3 minigene and FLAG-tagged Fox proteins were expressed in HEK293 cells as indicated and then subjected to the splicing assay described in Experimental Procedures. Alternative splicing products are depicted on the left of the gel. Error bars indicate SD (Right). (C) Minigene splicing assay with wild-type or UGCAUG element-mutated templates in the absence or presence of F:Fox-1 expression (mean ± SD). (D) Minigene splicing assay with F:Fox-1 and Myc-tagged proteins as indicated. Error bars show the SD in three experiments. (E) Endogenous FMNL3 splicing assay. Fox-2 down-regulation by siRNA against Fox-2 (SiFox-2) and F:Fox-1 expression were monitored by immunoblotting. (F) Effect of Myc-tagged WNK proteins on endogenous Fox-2–mediated FMNL3 splicing (mean ± SD).