Figure 3.
teNK cells promote colony formation.(A) Schema for teNK cell-tumor organoid coculture. (A′) Boxplot of tumor organoid invasion strands of tumor organoids cultured alone or in coculture with teNK cells. Error bars represent 5th to 95th percentile. ns, not significant by Mann–Whitney U test. (B) Schema for teNK cell-tumor cluster coculture. (B′) Normalized colony count of tumor clusters cultured alone or in coculture with teNK cells. Mean with SEM is represented. **, P < 0.01 by Mann–Whitney U test. (B″) Normalized colony count of tumor clusters cultured alone or in coculture with tiNK cells. Mean with SEM is represented. *, P < 0.05 by Mann–Whitney U test. (C) Schema for generating ceNK cells. (C′) Normalized colony count of MMTV-PyMT tumor clusters cultured alone or in coculture with ceNK cells. Mean with SEM is represented. **, P < 0.01 by Mann–Whitney U test. (D) Normalized colony count of MCF-7 cell clusters cultured alone or in coculture with ceHuNK cells. Mean with SEM is represented. **, P < 0.01 by Mann–Whitney U test. (E) Schema of the adoptive transfer of NK cells following a tail vein injection of cancer cells. (E′) Representative whole-lung images. Macrometastases were identified based on their mTomato expression. Scale bar, 4 mm. (E″) Boxplot of the number of lung macrometastases. Error bars represent 5th to 95th percentile. *, P < 0.05; ****, P < 0.0001 by Kruskal–Wallis test. (F) Heat map displaying z-scores for the variance-stabilized transform of gene expression for differentially expressed genes with absolute value of log2(fold change) >1 between hNK cells and teNK cells. Hierarchical clustering was used to order the genes. (G) Waterfall plot of genes associated with an active and resting NK cell phenotype, expressed by teNK cells and hNK cells. (H) Gene ontology enrichment analysis in “biological process” category for differentially expressed genes up- and down-regulated by teNK cells. Four categories with the lowest P value related to the immune system, metabolic processes, apoptosis, and proliferation are displayed.

teNK cells promote colony formation. (A) Schema for teNK cell-tumor organoid coculture. (A′) Boxplot of tumor organoid invasion strands of tumor organoids cultured alone or in coculture with teNK cells. Error bars represent 5th to 95th percentile. ns, not significant by Mann–Whitney U test. (B) Schema for teNK cell-tumor cluster coculture. (B′) Normalized colony count of tumor clusters cultured alone or in coculture with teNK cells. Mean with SEM is represented. **, P < 0.01 by Mann–Whitney U test. (B″) Normalized colony count of tumor clusters cultured alone or in coculture with tiNK cells. Mean with SEM is represented. *, P < 0.05 by Mann–Whitney U test. (C) Schema for generating ceNK cells. (C′) Normalized colony count of MMTV-PyMT tumor clusters cultured alone or in coculture with ceNK cells. Mean with SEM is represented. **, P < 0.01 by Mann–Whitney U test. (D) Normalized colony count of MCF-7 cell clusters cultured alone or in coculture with ceHuNK cells. Mean with SEM is represented. **, P < 0.01 by Mann–Whitney U test. (E) Schema of the adoptive transfer of NK cells following a tail vein injection of cancer cells. (E′) Representative whole-lung images. Macrometastases were identified based on their mTomato expression. Scale bar, 4 mm. (E″) Boxplot of the number of lung macrometastases. Error bars represent 5th to 95th percentile. *, P < 0.05; ****, P < 0.0001 by Kruskal–Wallis test. (F) Heat map displaying z-scores for the variance-stabilized transform of gene expression for differentially expressed genes with absolute value of log2(fold change) >1 between hNK cells and teNK cells. Hierarchical clustering was used to order the genes. (G) Waterfall plot of genes associated with an active and resting NK cell phenotype, expressed by teNK cells and hNK cells. (H) Gene ontology enrichment analysis in “biological process” category for differentially expressed genes up- and down-regulated by teNK cells. Four categories with the lowest P value related to the immune system, metabolic processes, apoptosis, and proliferation are displayed.

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