Suppression of metastases by RBM10 expression; validation of isoform-specific KDs. (A) Bioluminescence imaging of Luc+ 64860M cells 2 wk after TV injection showing decreased lung colonization upon Rbm10 expression. (B) Left: Bioluminescence imaging of Luc+ 64860M cells 3–4 wk after thyroid orthotopic implantation of 64860M cells ± Rbm10. Mice injected with Rbm10-expressing cells were sacrificed 8 days later to match tumor size with parental controls. Right: H&E stains of corresponding lung sections for each cohort. Arrows point to sites of lung metastatic colonization. (C–E) Validation of isoform-specific shRNA KDs by qRT-PCR using junction-spanning primers of the indicated exons in 64860M cells; (C) single, (D) dual, and (E) triple isoform KD in 64860M versus sh-Scramble demonstrating KD efficiency of the indicated AS isoforms (top) and the corresponding changes in total mRNA abundance (bottom). Data (C–E) represent mean with SD of four replicates. (F) Bioluminescence imaging and quantification 2 wk after TV injection of 64860M cells showing no changes in metastatic competence between sh.Scr and any of the individual isoform KDs. Statistical difference was analyzed using unpaired t test (C–F) and is indicated as ***P < 0.0001; ns: not significant. sh.Scr; sh-Scramble.