Supplementary Figure 1: Non-tumor breast tissue does not express TSLPR

a, Left panel: mRNA expression, examined by qPCR, of Tslp and other genes were tested in 4T1-M1 (non-targeting gRNA M1 in CRISPR-Cas9-GFP) and 4T1-Tslp^-/- 2-3 cells and were normalized to those in 4T1 cells to confirm the depletion of Tslp but intact of other genes in 4T1-Tslp^-/- 2-3 and no difference between 4T1 and 4T1-M1. n=3 per group. Right panel: TSLP protein expression in supernatant of 4T1-Tslp^-/- 2-3, 4T1-M1, and 4T1 cells in vitro culture for 2 days, examined by ELISA. n=3per group. b, Immunohistochemistry staining of TSLPR on adjacent non-tumor breast tissue (top) and on breast tumors collected (bottom) from breast cancer patients. A summary of breast cancer stage, tumor types, age, and gender from all recruited patients. Scale bar, 100 μm. Breast tumor tissue, n=12; adjacent non-tumor breast tissue, n=3. c, TSLPR and IL7R mRNA expression relative to expression of GAPDH, examined by qPCR, in MDA-MB-468 and MCF10A cells. n=3 per group. d, Left panel: mRNA expression of Tslpr and other genes in 4T1-Tslpr^-/- 4-6 and 4T1-M1 normalized to 4T1, examined by qPCR. n=3. Right panel: A representative histogram of surface TSLPR expression in 4T1-Tslpr^-/- 4–6, compared 4T1-M1 and 4T1 cells, examined by flow cytometry. IC: isotype control. Each symbol in (b) represents an individual patient and in (a, c, d left) represents individual cell culture. Data are represented as mean ± s.e.m. Statistical analysis by unpaired, two-tailed t test with 95 % confidence intervals. Results in (a, d) are representative of two independent experiments.