Figure 2.
hNK cells induce apoptosis in K14+ invasive breast cancer cells.(A) Representative confocal images of the invading strands of tumor organoids (mTomato+) and caspase activity (green) cultured alone (top) or in coculture with hNK cells (bottom). Scale bar, 10 µm. (A′ and A″) Boxplot of the percentage of organoids with caspase activity in invading strands per biological replicate (A′) and the total number of invading strands with caspase activity (A″) cultured alone or in coculture with hNK cells. *, P < 0.05; ****, P < 0.0001 by Mann–Whitney U test. (B) Representative confocal images of tumor clusters organoids (mTomato+) and caspase activity (green) cultured alone (top) or in coculture with hNK cells (bottom). Scale bar, 10 µm. (B′) Boxplot of the percentage of tumor clusters per biological replicate with caspase activity in tumor clusters cultured alone or in coculture with hNK cells. *, P < 0.05 by Mann–Whitney U test. (C) Schema for assessing IFNγ activity in hNK cells in response to coculture with K14+ or K14− tumor cells. hNK cells were taken from ROSAmT/mG mice and in coculture with K14+ or K14− cells from K14-actin-GFP;MMTV-PyMT mice. In this experiment, hNK cells were fluorescently labeled with mTomato and K14+ cells were labeled with GFP. (C′) Boxplot of IFNγ expression among hNK cells after coculture with K14+ and K14− cells and normalized to K14− cells. Error bars represent 5th to 95th percentile. ***, P < 0.001 by Mann–Whitney U test. (D) Schema for assessment of the innate immune response to an initial metastatic seed. Tumor clusters from the mammary tumors of K14-actin-GFP;MMTV-PyMT;ROSAmT/mG mice were injected into the tail veins of immunocompetent mice, and the lung microenvironment was assessed after 6 h. (D′) Boxplot of the number of NK cells, macrophages, and neutrophils around a metastatic seed. Error bars represent 5th to 95th percentile. ***, P < 0.001; ****, P < 0.0001 by Kruskal–Wallis test. (D″) Representative slide scanned images of lung tissue field of view containing a K14+ (green), metastatic seed (magenta), surrounded by NK cells (NK1.1, white). Scale bar, 20 µm.

hNK cells induce apoptosis in K14+ invasive breast cancer cells. (A) Representative confocal images of the invading strands of tumor organoids (mTomato+) and caspase activity (green) cultured alone (top) or in coculture with hNK cells (bottom). Scale bar, 10 µm. (A′ and A″) Boxplot of the percentage of organoids with caspase activity in invading strands per biological replicate (A′) and the total number of invading strands with caspase activity (A″) cultured alone or in coculture with hNK cells. *, P < 0.05; ****, P < 0.0001 by Mann–Whitney U test. (B) Representative confocal images of tumor clusters organoids (mTomato+) and caspase activity (green) cultured alone (top) or in coculture with hNK cells (bottom). Scale bar, 10 µm. (B′) Boxplot of the percentage of tumor clusters per biological replicate with caspase activity in tumor clusters cultured alone or in coculture with hNK cells. *, P < 0.05 by Mann–Whitney U test. (C) Schema for assessing IFNγ activity in hNK cells in response to coculture with K14+ or K14− tumor cells. hNK cells were taken from ROSAmT/mG mice and in coculture with K14+ or K14− cells from K14-actin-GFP;MMTV-PyMT mice. In this experiment, hNK cells were fluorescently labeled with mTomato and K14+ cells were labeled with GFP. (C′) Boxplot of IFNγ expression among hNK cells after coculture with K14+ and K14− cells and normalized to K14− cells. Error bars represent 5th to 95th percentile. ***, P < 0.001 by Mann–Whitney U test. (D) Schema for assessment of the innate immune response to an initial metastatic seed. Tumor clusters from the mammary tumors of K14-actin-GFP;MMTV-PyMT;ROSAmT/mG mice were injected into the tail veins of immunocompetent mice, and the lung microenvironment was assessed after 6 h. (D′) Boxplot of the number of NK cells, macrophages, and neutrophils around a metastatic seed. Error bars represent 5th to 95th percentile. ***, P < 0.001; ****, P < 0.0001 by Kruskal–Wallis test. (D″) Representative slide scanned images of lung tissue field of view containing a K14+ (green), metastatic seed (magenta), surrounded by NK cells (NK1.1, white). Scale bar, 20 µm.

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