Impact of RBM10-induced AS of VCL, CD44, and TNC on cell motility and invasiveness. (A and B) IPA of RNAseq of RBM10-null versus RBM10-expressing PE121410 cells showing top 25 functional annotations (A) and canonical pathways (B). Highlighted in red are pathways involved in cell migration, movement, invasiveness, RHO-RAC signaling, and ECM–cytoskeleton interactions. (C) Western blotting for RBM10 and β-actin in the indicated cell lines. (D) Top two rows: Representative cell trajectories by time-lapse imaging of RBM10-null KTC1 and PE121410 at baseline and after dox induction of RBM10 for 48 h. Third row: Cell trajectory of 8305C cells transduced with scrambled or RBM10 shRNAs. (E) Mean cell velocity quantified by time-lapse imaging for the three cell lines ± RBM10. (F) Western blot (left), RT-PCR (middle), and qRT-PCR (right) for VCL, CD44, and TNC, respectively, in KTC1 cells transduced with sh.Renilla or shRNAs targeting the indicated exons demonstrating isoform-specific KDs. qRT-PCR data represent mean with SEM of four replicates. (G–I) Cell migration velocity by time-lapse imaging (left) and cell invasion by transwell assay (right) in control versus isoform-specific KD of VCL (G), CD44 (H), and TNC (I). (J and K) Increased RAC1-GTP levels (J) and RAC1 downstream signaling (pPAK and pAKT-S473) (K) in RBM10-null PE121410 and KTC1 cells. Statistical difference was analyzed using unpaired t test (E, F—right, and G–I) and is indicated as *P < 0.05, **P < 0.01, and ***P < 0.0001; ns: not significant. IPA: Ingenuity Pathway Analysis. Source data are available for this figure: SourceData F3.