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. Author manuscript; available in PMC: 2015 Oct 23.
Published in final edited form as: Cell. 2014 Oct 23;159(3):499–513. doi: 10.1016/j.cell.2014.09.051

Figure 1. Stromal cells induce ISGs and protect basal-like breast cancer cells against radiation in a STAT1-dependent manner.

Figure 1

A) Human MDA-MB-231 metastatic breast cancer cell (BrCa) line (1833) was admixed with or without MRC5 normal human fibroblasts (Stroma) and expression of IRDS genes was determined by qRT-PCR. B) GFP-labeled 1833 breast cancer cells with and without MRC5 fibroblasts were xenografted subcutaneously into nude mice and tumors imaged (20X) at day 14. STAT1 intensity in breast cancer cells is quantitated for representative field shown. Scale bar is 100 microns. C) Bioluminescence imaging (BLI) response of 1833 breast cancer cells with a luciferase reporter gene after xenografting with and without MRC5 fibroblasts. Tumors were irradiated with 8 Gy (day 0). Shown is change in photon flux over time (first derivative, mean ± SEM, n=5–10). Positive first derivative indicates growth, zero indicates no growth, and negative values denote regression. Data are a separate analysis of the control groups from Figure 5M. D) 1833 breast cancer cells were stained with GFP and TUNEL (red) 10 days after RT. Percent TUNEL positive is shown. Scale bar is 100 microns. E) Breast cancer cells (Table S1) were classified as IRDS responders (IRDS-Rs) or IRDS non-responders (IRDS-NRs). Heat map and scale shows breast cancer IRDS genes after mono-culture (M) or MRC5 co-culture (C). F) Cell death of IRDS-Rs and IRDS-NRs four days after 10 Gy RT in mono- (Mono) and co-culture (Co-cx) (n=3–10). G) Cell death of 1833 IRDS-R after cisplatin chemotherapy (n=3) and after dose response. H) Gene Set Analysis shows changes in IRDS genes 48 hrs after co-culture vs mono-culture of IRDS-Rs (left, also see Table S1), or after STAT1 knockdown in 1833 IRDS-R in co-culture (right). Top graph plots individual and overall gene scores, and bottom graph shows fold-change. I) Cell death of 1833 IRDS-R four days after 10 Gy RT using three independent siRNAs to STAT1. J) BLI-based survival assay after 10 Gy RT (day 0) using luciferase-labeled 1833 cells with shSTAT1 or control knockdown (shCont). Photon flux (×106) for each well is indicated. Shown is representative experiment (n=5). *p < 0.05. See Figure S1.