Figure 3.

Ccl5 deletion in lung cancer cells reduces tumor burden. LLC cells were infected with lentiviral particles containing CRISPR/cas9 guide targeting Ccl5 (sgCCL5) or a scramble sequence (sgCNTRL). (a) CCL5 production was quantified by ELISA. Data show n = 3, significance by students t-test, bars indicate mean ± SEM. (b) Ccl5 deletion did not alter the in vitro growth rate of LLC cells determined by real-time cell confluence tracking by Incucyte as % confluency over time. (c) Day 14 after immune deficient NRG mice injected i.v. with Ccl5 deletion cells did not display a significant difference in lung tumor burden. Data show n = 8-9 , bars indicate mean ± SEM. (d) Day 19 after immune competent C57BL/6 mice injected i.v. with Ccl5 deletion cells had reduced tumor burden in the lungs. Data show n = 8, bars indicate mean ± SEM. Tumor burden was quantified as the fraction of tumor area/total lung area from 5 step sections 150 µm apart. Scale bars represent 2000-5000 µm. Significance from Graphpad Prism was determined using an ANOVA with a post-hoc Tukey’s test. If data did not have normal distribution a Kruskal-Wallis test was performed. *P < .05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001