Figure 6: GALNT14 enables BCCs to modify and exploit the lung microenvironment for their metastatic outgrowth. | Nature Communications

Figure 6: GALNT14 enables BCCs to modify and exploit the lung microenvironment for their metastatic outgrowth.

From: GALNT14 promotes lung-specific breast cancer metastasis by modulating self-renewal and interaction with the lung microenvironment

Figure 6

(a) Immunofluorescent staining of frozen lung sections from mice injected with the indicated 231-LM2 cells by using an antibody against a macrophage marker F4/80. Representative images from at least three independent lung samples are shown. F4/80 (magenta), GFP (cyan), DAPI (blue). GFP-positive cells represent cancer cells. Scale bar, 100 μm. (b) Relative RAW 264.7 cell transwell migration by CM from the indicated 231-LM2 (left) and 231-Par cells (right). n=6. (c) Similar experiments as in b except that CM were collected from shCntr and shGALNT14-expressing 231-LM2 cells transfected with scrambled (siScr) and two independent siRNAs against CXCL1. n=9. (d) The role of GALNT14 in macrophage-stimulated growth of BCCs. The indicated 231-LM2 cells were cultured alone or co-cultured with RAW 264.7 cells but separated by a permeable membrane. Cell lysates were then subjected to immunoblotting analysis by using antibodies against the indicated proteins. Western blot images are representative of three independent experiments. (e,f) The indicated 231-LM2 cells were co-cultured in contact with (e) or separated from RAW 264.7 cells by a permeable membrane (f) for 3 days. The growth of 231-LM2 cells in the presence of RAW 264.7 was measured by luminescent signals and normalized to that in the absence of RAW 264.7. n=6 (e) and 9 (f). (g,h) Paraffin-embedded lung sections from mice injected with the indicated 231-LM2 (g) and 231-Par (h) cells were subjected to the immunohistochemistry with an antibody against Ki-67. Left: Quantification of Ki-67 staining. n=9. Right: Representative Images. Scale bar, 50 μm. P values were calculated using two-tailed unpaired Student’s t-test. Results represent mean±s.e.m. (error bars).

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