Journal: Cell Death & Disease

Article Title: Effects of methionine deficiency on B7H3-DAP12-CAR-T cells in the treatment of lung squamous cell carcinoma

doi: 10.1038/s41419-023-06376-w

Figure Lengend Snippet: A Expression of CD276 in LUSC ( n = 502) and normal samples ( n = 51) based on TCGA. B The relationship between CD276 and CD8 + T cells (R = −0.188, P < 0.001) or CD4+ activated memory cells (R = −0.138, P < 0.001) in LUSC analyzed by Timer2.0. C Histochemistry score (H-score) of B7H3 on 10 samples of normal lung tissues, LUAD, LUSC, and SCLC shown by IHC staining. Scale bars = 200 μm. D H-score of B7H3 on 48 pairs of LUSC tissues and adjacent normal tissues shown by IHC. Scale bars = 100 μm. E CD276 expressed in SK-MES-1, H520, H1703, and B2B shown by qRT-PCR. F B7H3 localization in H1703/H520/SK-MES-1 shown by IF (×40 magnification). Representative images are shown. Scale bars = 100 μm. G B7H3 expression on the cell membrane of H1703 and H520 shown by flow cytometry. * P < 0.05; ** P < 0.01; *** P < 0.001. ns, not significant. Variables are presented as mean ± SD.

Article Snippet: LUSC cell lines (H1703, H520, and SK-MES-1) and the pulmonary epithelial cell BEAS-2B (B2B) were purchased from China Center Type Culture Collection (CCTCC, Shanghai) and cultured in RPMI-1640 (GIBCO, USA) supplemented with 10% FBS, 100 U/ml penicillin and 100 mg/ml streptomycin (Invitrogen, USA) at 37°C and 5% CO2.

Techniques: Expressing, Immunohistochemistry, Quantitative RT-PCR, Membrane, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Effects of methionine deficiency on B7H3-DAP12-CAR-T cells in the treatment of lung squamous cell carcinoma

doi: 10.1038/s41419-023-06376-w

Figure Lengend Snippet: A Normalized cell index of B7H3-DAP12-CAR-T co-cultured with target cells at different ratios (E:T = 0:1, 1:1, 2:1, 5:1) in 25 or 100 μM Met within 28 h. B Normalized cell index of LUSC cell lines at 10, 25, 50, 75, or 100 μM Met within 24 h. C Normalized cell index of B7H3-DAP12-CAR-T co-cultured with target cells (E:T = 0:1 or 2:1) at 25, 50, 75, or 100 μM Met. D IL-2 and IFN-γ secreted by B7H3-DAP12-CAR-T co-cultured with target tumor cells (2 × 10 5 ) at E:T = 2:1 for 48 h in 25, 50, 75, or 100 μM Met measured by ELISA. E Expression of cytokine-associated genes in CAR-T co-cultured with 1703-B7H3 (E:T = 1:1) at 25 or 100 Met shown by qRT-PCR. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant. Variables are presented as mean ± SD.

Article Snippet: LUSC cell lines (H1703, H520, and SK-MES-1) and the pulmonary epithelial cell BEAS-2B (B2B) were purchased from China Center Type Culture Collection (CCTCC, Shanghai) and cultured in RPMI-1640 (GIBCO, USA) supplemented with 10% FBS, 100 U/ml penicillin and 100 mg/ml streptomycin (Invitrogen, USA) at 37°C and 5% CO2.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: Effects of methionine deficiency on B7H3-DAP12-CAR-T cells in the treatment of lung squamous cell carcinoma

doi: 10.1038/s41419-023-06376-w

Figure Lengend Snippet: A Expression of classical Met transporters (SLC7A5, SLC7A6, SLC7A7, SLC7A8,SLC38A1, SLC38A2, SLC43A2) in LUSC ( n = 502) relative to normal adjacent tissues ( n = 51) based on TCGA shown by the heatmap and bar chart. B Expression of SLC7A5 among 26 types of LUSC cell lines based on DepMap. C A Venn diagram consisting of SLCs highly expressed in LUSC based on TCGA, SLCs highly expressed in LUSC cell lines based on DepMap, and SLCs negatively associated with the overall survival of LUSC patients based on TCGA. D The relationship between SLC7A5 and CD8 + T cells (R = –0.224, P < 0.001) in LUSC analyzed by Timer2.0. E SLC7A5 expressed in SK-MES-1, H520, H1703, and B2B shown by qRT-PCR. F SLC7A5 expressed in SK-MES-1, H520, H1703, and B2B shown by WB (left); SLC7A5 expression in 1703-B7H3 and CAR-T after co-cultured (E:T = 2:1) for 48 h in 25 or 100 μM shown by WB (right). G After culturing for 2 days, Met concentration in supernatant of 1 × 10 5 LUSC cells with SLC7A5 up- or down-regulated detected by ELISA. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant. Variables are presented as mean ± SD.

Article Snippet: LUSC cell lines (H1703, H520, and SK-MES-1) and the pulmonary epithelial cell BEAS-2B (B2B) were purchased from China Center Type Culture Collection (CCTCC, Shanghai) and cultured in RPMI-1640 (GIBCO, USA) supplemented with 10% FBS, 100 U/ml penicillin and 100 mg/ml streptomycin (Invitrogen, USA) at 37°C and 5% CO2.

Techniques: Expressing, Quantitative RT-PCR, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay