Fig. 2. SERPINB9 regulates T-cell-mediated killing and lung tumor growth.
A Growth curves of tumors in KrasG12D;Cas9 mice transduced with sgNTC or sgSerpinb9 untreated or ACT treated. (CTRL: untreated n = 10; treated, n = 10 for each group) The p values for tumor growth were determined by an unpaired two-sided t-test with Welch’s correction. The data are presented as the mean ± SEM. B Growth curves of tumors in BrafV600E;Cas9 mice transduced with sgNTC or sgSerpinb9 untreated or ACT treated. (CTRL: untreated n = 7 and treated n = 9; sgSerpinb9: untreated n = 6 and treated n = 6). The p values for tumor growth were determined by an unpaired two-sided t-test with Welch’s correction. The data are presented as the mean ± SEM C Tumor-free survival of KrasG12D;Cas9 mice from (A). D Tumor-free survival of Braf V600E;Cas9 mice from (B). Comparison of survival curves was performed by Log-rank (Mantel–Cox test). E The effect of Serpinb9 knockout on T-cell-mediated tumor killing. LLC cells transduced with sgNTC or sgSerpinb9 and labeled with CFSE were used as targets (T) and cocultured with activated OT-I effector cells (E) at different E:T ratios for 4 h. Flow cytometry analysis was used to quantify the percentage of target cells killing at different E:T ratios. Data presents mean ± s.e.m. of three technical replicates analyzed by multiple unpaired t-tests. One representative experiment out of three independent experiments is shown. F, G The effect of SERPINB9 overexpression on DNT cell-mediated killing. Human LUAD cell lines, A549 and H125 labeled with CFSE were used as targets (T) and cocultured with activated and expanded DNT effector cells (E) from two different donors (UPN119, UPN133) at different E:T ratios for 18 h. Data presents mean ± s.e.m. of three technical replicates analyzed by multiple unpaired t-tests. One representative experiment out of two independent experiments is shown for each donor.