Effect of SRSF1 and SRSF3 on alternative splicing of Rac1b in NCM460 cells. (A) The normal colonocyte cell line NCM460 was cotransfected with a RAC1 minigene and the indicated T7-tagged splicing factors. The Western blot detection of T7-SRSF1 and T7-SRSF3 is shown 24 h after transfection in NCM460 with endogenous α-tubulin as loading control. qPCR quantification of the corresponding minigene-derived transcripts is shown in the lower panel. (B) NCM460 cells were transfected with the indicated siRNAs and analyzed after 48 h. A Western blot detection of endogenous levels of Rac1b is shown, together with those of SRSF1 and SRSF3, to document successful knockdown and of α-tubulin as loading control. The lower panel shows quantification of the corresponding splicing changes in endogenous Rac1 and Rac1b transcript levels by qPCR.
