Supplementary Figure S15 from Stress Promotes Lung Metastasis in Breast Cancer by Altering Neutrophil Differentiation

Conditional knockout of Ccl3/Ccl4 in neutrophils using an optimized CRISPR/Cas9 approach. A. Schematic representation of the main functional elements in the CRISPR plasmid designed to specially target Ccl3 and Ccl4 in neutrophils. pSpCas9(BB)-2A-Puro (PX459) was used as the vector backbone. B. Sequences of Guide RNAs (gRNAs) designed to target the mouse Ccl3 and Ccl4 genes. C. Western blot analysis of CCL3 and CCL4 in primary neutrophils from the lungs of breast tumor-burden mice in both NC and CS groups with or without knockout of Ccl3/Ccl4.