Fig 2: PEM-induced immunogenicity is restrained by CD73. A, Tumor volume of HCC827GR6 cells with Scramble (Scr) or CD73sgRNA using humanized murine model (n = 8 in each group). B, Frequency of CD8+ T cells infiltration among CD45+ cells in mouse tumors (n = 5 in each group). C and D, Representative CD8 IHC images and analysis from Scramble or CD73sgRNA mouse tumors. Red arrows highlight intratumoral localization of CD8+ T cells. Scale bar, 100 µmol/L. E, Schematic of study with PEM in humanized murine HCC827GR6 cells models. F, Tumor volume of HCC827GR6 cells with Scramble or CD73 sgRNA with PEM treatment in humanized murine model (n = 3 in each group). G and H, HLA-A,B,C expression on HCC827, HCC827GR6, and HCC827EPR cells treated ± PEM for 48 hours, followed by 24 hours of drug-free. MFI, mean fluorescence intensity. ?MFI, (Pemetrexed-DMSO (dimethyl sulfoxide))/DMSO. Mean ± SEM of n = 5 biological replicates is shown (unpaired two-tailed Student t test). I, CD73 expression on HCC827GR6 cells treated ± PEM for 48 hours, followed by 24 hours of drug-free. J, Adenosine levels in CM derived from HCC827GR6 cell lines with Scramble or CD73 sgRNA treated ± PEM for 48 hours for 72 hours. Mean ± SEM of n = 3 biological replicates is shown (unpaired two-tailed Student t test). K, ELISA of human IFN? and IL2 levels in CM derived from HCC827GR6 cell lines with Scramble or CD73 sgRNA pretreated ± PEM for 48 hours with J76 cells transduced with LMP2B TCR for 72 hours and pretreated ±1 µg/mL peptides (SSCSSCPLSK) for 3 hours. Mean ± SEM of n = 3 biological replicates is shown (unpaired two-tailed Student t test). L, Schematic of treatment strategy on in MET-driven EGFR-TKI–resistant cells. *, P < 0.05; **, P < 0.005; ***, P < 0.001; n.s., nonsignificant.