FIGURE 5.

RBM15B inhibits in vivo splicing of β-galactosidase/luciferase reporter system. Expression constructs for RBM15B WT, N-term, ΔSPOC, Core, and Core/NLS and empty vector were transiently co-transfected into HEK293T cells with the pTN24 splicing reporter plasmid for 24 h. RBM15B expression vector was titered to determine the optimal amount for in vivo splicing assays (0.5 μg). A, relative splicing activities were calculated using the ratios of luciferase to β-galactosidase activities with the activity of the control cells transfected with empty expression vectors (Control Vector) set at 1. The results shown represent three to six independent transfection experiments in which one plate was transfected, and each lysate was measured in quadruplicate. The error bars represent the S.E. and (p) denotes probability values. * denotes p < 0.001 compared with empty vector calculated using a non-parametric rank model. B, cell lysates were analyzed by immunoblotting for expression of RBM15B proteins (anti-RBM15B antibody). Relative protein loading was confirmed by immunoblotting with anti-actin antibody.