Supplementary Methods, Figures 1-9 from CCL2 Mediates Cross-talk between Cancer Cells and Stromal Fibroblasts That Regulates Breast Cancer Stem Cells

PDF file - 111K, Methods: Includes Mammosphere formation assay, Cell transfection and reporter assays, RNAi studies, as well as Western blot analyses and xenograft data. Figures S1-S9 include the following: Marker expression in primary fibroblasts and BC cells (S1); Serial confocal images of representative mammospheres formed by PKH67-labeled BT474 cells in the presence or absence of CCL2 (S2); CCL2 expression in CAF3 cells upon CM treatments (S3); Sustained CCL2 induction in CAFs co-cultured with tumor cells (S4); Expression of CCR2 and CCR4 in BC cells (S5); HES1 and HEY1 reporter assays in BT474 cells (S6); In vitro secretase cleavage assay (S7); Representative IHC images of XP/CAF265922 xenograft tumors treated with PBS, IgG, or anti-CCL2 (S8); Representative IHC images of CCL2 and NOTCH1 in primary BC specimens (S9).