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Fig. 2 | Molecular Cancer

Fig. 2

From: N6-methyladenosine-modified circIGF2BP3 inhibits CD8+ T-cell responses to facilitate tumor immune evasion by promoting the deubiquitination of PD-L1 in non-small cell lung cancer

Fig. 2

circIGF2BP3 is overexpressed in NSCLC and attenuates CD8+ T cell-mediated antitumor immunity. A The relative expression of circIGF2BP3 in 68 paired samples of NSCLC in cohort I from Tongji Hospital. GAPDH was used as an internal control. B Kaplan-Meier (K-M) plot showing the relationship between circIGF2BP3 levels and patient OS in cohort I from Tongji Hospital. C Cox regression analysis showed that circIGF2BP3 was an independent prognostic biomarker in NSCLC. D Representative images and statistical quantitation of FACS analysis of PBMC-mediated elimination of NSCLC cells, as determined by annexin V-FITC and propidium iodide (PI) double labeling. E Expression of perforin and GzmB in PBMCs after coculture with circIGF2BP3-knockdown A549 cells or circIGF2BP3-overexpressing SK-MES-1 cells. F The levels of IFN-γ and TNF-α secreted by PBMCs were detected by ELISA after coculturing with circIGF2BP3-knockdown A549 cells or circIGF2BP3-overexpressing SK-MES-1 cells. G Ki-67 staining of PBMCs after coculturing with circIGF2BP3-OE or circIGF2BP3-silenced NSCLC cells. Scale bars, 20 μm. H TUNEL staining of PBMCs after coculturing with circIGF2BP3-OE or circIGF2BP3-silenced NSCLC cells. Scale bars, 20 μm. I Schematic diagram exhibiting the grouping and treatment plan of the syngeneic mouse model: C57BL/6 mice were inoculated with 106 CTRL or circIGF2BP3-OE LLC cells and received CD8 mAb treatment or IgG2b control treatment at the indicated time points. J Time-course evaluation of tumor volumes measured every 3 days. K Representative flow cytometry data for infiltrated CD8+ T cells in CTRL and circIGF2BP3-OE LLC xenografts. L-M Representative images and statistical quantitation of the FACS analysis of the percentage of IFN-γ+, TNF-α+, GzmB+ and perforin+ CD8+ TILs from CTRL and circIGF2BP3-OE LLC xenografts. Data represent the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. P values were determined by unpaired Student’s t test (F, G and H), paired Student’s t test (A), one-way ANOVA with Tukey’s post hoc test (D, F, G, H, J, K, L and M) and log-rank test (B)

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Molecular Cancer

ISSN: 1476-4598