Fig. 6From: Predictive biomarkers for the responsiveness of recurrent glioblastomas to activated killer cell immunotherapy Effects of three candidate genes on migration of NK cells toward GBM cancer cells. AÂ Expression of TNFSF4, TNFRSF18, and IL12RB2 in T cells and U87MG cells was analyzed by flow cytometry. Left: Representative histogram plots. Gray histograms represent isotype controls. Right: Column graphs show the frequencies of TNFSF4+, TNFRSF18+, and IL12RB2+ cells. BÂ Transwell migration assay: CFSE-stained NK cells were plated in the upper chamber of a Transwell insert with an 8-mm pore size. Cells were allowed to migrate for 8Â h toward U87MG cells with or without T cells in the lower chamber. T cells were pre-incubated with blocking antibodies to TNFSF4, TNFRSF18, and IL12RB2 for 1Â h. NK cells that migrated to the lower chamber were counted using a cell counter. The experiments were performed in triplicateBack to article page