Figure 1 from Epigenetic Induction of Cancer-Testis Antigens and Endogenous Retroviruses at Single-Cell Level Enhances Immune Recognition and Response in Glioma

CpG island methylation is associated with CTAG1A/B downregulation in the majority of gliomas and DAC time-dependently induces NY-ESO-1 expression via promoter demethylation for targeting by NY-ESO-1-specific TCR.A, Representative lollipop diagram of direct methylation PCR sequencing of immortalized GBM cell lines (U-251MG, BT142), immortalized meningioma cell line (CH-157MN), primary GBM cell lines (HK211, HK213, HK308, HK252), IDHwt (n = 17) and IDHmut (n = 10) patient samples. M, Methylated and U, Unmethylated determined from cytosine/thymine (C/T) peak ratio. B, Representative Western blot of native CH-157MN and U-251MG and U-251MG treated with 1 µmol/L DAC for 2 days. C, Cytotoxic analysis of U-251MG and CH-157MN cells over a 40-hour period when co-incubated with NY-ESO-1 TCR-transduced effector T cells (n = 2; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; unpaired t test). D, Representative Bis-Seq-TA cloning lollipop diagram and (E) percent methylated CpGs to total CpGs of first eight sites of analyzed CpG island in U-251MG cells treated with DMSO or 1-µmol/L DAC for 2 or 4 days. F, Average fold change of mRNA expression of treated U-251MG (n = 3; *, P < 0.05; one-way ANOVA). G, Cytotoxic analysis of treated U-251MG cells over a 40-hour period when co-incubated with NY-ESO-1 TCR-transduced effector T cells at E:T ratio of 1:1 and (H) at multiple E:T ratios (1:10, 1:5, 1:2, 1:1) at 20-hour (n = 3; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; multiple unpaired t tests).