FIG 5.

Combinatorial regulation of AS during EMT by ESRP1/2 and RBM47. (A) Validation by qRT-PCR of ESRP1, ESRP2, and RBM47 mRNA depletion using siRNAs (in biological triplicates). (B) Western blot validation of knockdown of ESRP1/2 and RBM47 protein. Note that RBM47 mRNA showed a 50% reduction upon ESRP1/2 knockdown, while the protein level did not change appreciably. (C) Validation of representative exons where ESRP1/2 and RBM47 knockdown have additive functions to promote splicing changes that occur during EMT. p1, p2, and p3 are P values comparing siESRP1/2, siRBM47, and siCombined to siCtrl, respectively, and p4 is to compare day 7 to no Dox. (D) Validation of representative exons where ESRP1/2 and RBM47 promote different or opposite changes in splicing. For MYO1B, there are two consecutive cassette exons (both are 87 nt in length) that can be included individually (middle band) or together in tandem (top band) or skipped together (bottom band). Only the top and bottom bands are quantified to calculate the PSI.