Journal: Canadian Journal of Gastroenterology & Hepatology

Article Title: Depletion and Reversal of Hepatocellular Carcinoma Inducing CTL through ER Stress-Dependent PERK-CHOP Signaling Pathway

doi: 10.1155/2022/6413783

Figure Lengend Snippet: Function of CTLs was induced by hepatoma cells. (a) CTLL-2 cells were cocultured with hepatoma cells. PD-1 and TIM-3 expressions were determined by western blotting analysis. (b) PRF1 and GzmB expression was analyzed by ELISA. Data are presented as the mean ± SD. ∗∗∗ P < 0.001, ns indicates no statistical significance.

Article Snippet: The supernatant was aliquoted into a centrifuged tube and used for the concentration-dependent detection of PRF1 (E-EL-M0890c, ElabScience, China) and GzmB (ELMO003, Boxbio, China) in strict accordance with the manufacturer's instructions.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Canadian Journal of Gastroenterology & Hepatology

Article Title: Depletion and Reversal of Hepatocellular Carcinoma Inducing CTL through ER Stress-Dependent PERK-CHOP Signaling Pathway

doi: 10.1155/2022/6413783

Figure Lengend Snippet: Activation of ER stress induces CTLs depletion. (a) CTTL-2 cells were cocultured with Hepa1-6 cells for one day. Different concentrations of TM (0, 1, 2, and 4 μ M) were added to CTTL-2 cells and cultured for 1 day. Western blotting analysis was used to determine GRP78 levels. (b) Cells were treated with TM (0 and 4 μ M), and changes in ER morphology were analyzed by electron microscopy (scale bar = 2 μ m). (c) CTTL-2 cells were cocultured with Hepa1-6 cells for one day. Different concentrations of 4-PBA (0, 0.1, 0.2 and 0.4 mM) were added to CTTL-2 cells and cultured for one day. The expression of GRP78 was analyzed by western blotting. (d) Cells were treated with 4-PBA (0 and 0.4 mM), and ER morphology was examined by electron microscopy. (e) CTLL-2 cells were treated with TM, and western blotting was used to analyze PD-1 and TIM-3 expression. (f) ELISA analysis of PRF1 and GzmB expression. (g) CTLL-2 cells were treated with 4-PBA, and PD-1 and TIM-3 expression levels were measured by western blotting. (h) ELISA was used to detect the expression of PRF1 and GzmB. Data are presented as the mean ± SD. ∗∗∗ P < 0.001, ns indicates no statistical significance.

Article Snippet: The supernatant was aliquoted into a centrifuged tube and used for the concentration-dependent detection of PRF1 (E-EL-M0890c, ElabScience, China) and GzmB (ELMO003, Boxbio, China) in strict accordance with the manufacturer's instructions.

Techniques: Activation Assay, Cell Culture, Western Blot, Electron Microscopy, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Canadian Journal of Gastroenterology & Hepatology

Article Title: Depletion and Reversal of Hepatocellular Carcinoma Inducing CTL through ER Stress-Dependent PERK-CHOP Signaling Pathway

doi: 10.1155/2022/6413783

Figure Lengend Snippet: CTLs depletion was induced through the ER stress PERK-CHOP pathway. After coculturing hepatoma cells with CTLL-2 cells, (a) western blotting was used to examine the expression of PERK, P-PERK, and CHOP. (b) TM-induced protein expression of PERK, P-PERK, and CHOP. (c) 4-PBA treatment induced a reduction in PERK, P-PERK, and CHOP protein levels. (d) Cells were cocultured for one day and CTLL-2 cells were treated with either GSK2656157 (0, 10, 50 and 100 nM) for one day. Western blotting analysis of ER stress-associated protein expression. (e) Electron microscopic analysis (scale bar = 2 μ m). (f) Protein expression of PD-1 and TIM-3 in CTLL-2 cells. (g) Expression of PRF1 and GzmB in CTLL-2 cells. (h) Expression of ER stress-associated proteins in CTLL-2 cells after one day of silencing CHOP plasmid (0 and 0.25 μ g/ μ L) treatment. (i) ER morphology was detected by electron microscopy (scale bar = 2 μ m). (j) Western blotting and (k) ELISA analysis. Data are presented as the means ± SD. ∗∗∗ P < 0.001, ns indicates no statistical significance.

Article Snippet: The supernatant was aliquoted into a centrifuged tube and used for the concentration-dependent detection of PRF1 (E-EL-M0890c, ElabScience, China) and GzmB (ELMO003, Boxbio, China) in strict accordance with the manufacturer's instructions.

Techniques: Western Blot, Expressing, Plasmid Preparation, Electron Microscopy, Enzyme-linked Immunosorbent Assay

Journal: Cancer Drug Resistance

Article Title: Effects of FAP + cancer-associated fibroblasts on anti-PD-1 immunotherapy and CD4 + T cell polarization in gastric cancer

doi: 10.20517/cdr.2025.97

Figure Lengend Snippet: FAP + CAFs may inhibit CD8 + T cell function by promoting the polarization of naive CD4 + T cells toward Th2. (A) Flow cytometric analysis of the effect of FAP + CAFs on CD8 + T cell proliferation with or without the presence of naive CD4 + T cells. (B and C) ELISA measuring whether IL-31 antibodies can block the decrease in IFN-γ and perforin expression in CD8 + T cells mediated by co-culture with FAP + CAFs and CD4 + T cells ( n = 3). ** P < 0.01; *** P < 0.001; NS: no significance; bars: means ± standard deviation. CAFs: Cancer-associated fibroblasts.

Article Snippet: Plasma and cell culture supernatant levels of IL-31 (Elabscience, E-EL-H5469), IFN-γ (Elabscience, E-EL-H0108), perforin (Elabscience, E-EL-H1123), and IL-4 (Elabscience, E-EL-H0101) were measured by ELISA kits according to the manufacturer’s instructions.

Techniques: Cell Function Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay, Expressing, Co-Culture Assay, Standard Deviation

Journal: Acta Pharmaceutica Sinica. B

Article Title: A triple combination strategy for nasopharyngeal carcinoma: Aptamer-guided liposomal chemotherapy, engineered NK cells, and Fc-enhanced PD-L1 antibody therapy

doi: 10.1016/j.apsb.2025.10.007

Figure Lengend Snippet: Aptamer-engineered NK cells exhibit enhanced targeted cytotoxicity and effector functions against NPC cells. (A) Cytotoxicity of NK, S3-NK, P-NK, and S3-P-NK cells against 5-8F target cells, measured by LDH release assay across a range of effector-to-target (E:T) ratios after a 2 h co-culture. (B) Flow cytometry images demonstrated that NP69, 5-8F, and C666-1 cells were co-cultured with NK cells at a 10:1 E:T ratio for 2 h, washed with PBS, and co-incubated for 24 h, leading to apoptosis and necrosis. (C) Quantification of total apoptotic (early + late) and necrotic cell populations from the analysis shown in (B). (D) Schematic of the proposed mechanism for S3-P-NK cell-mediated antitumor immunity, involving targeted recognition followed by the release of cytotoxic granules (perforin, granzyme B) and immunostimulatory cytokines (IFN- γ ). (E–G) ELISA quantification of effector molecules released into the supernatant after co-culture of NK cells with 5-8F or C666-1 target cells (E:T = 10:1) for 2 h, PBS washing, and co-incubation for 24 h: (E) IFN- γ , (F) Granzyme B, (G) Perforin. Data in (A, C, E, F, G) are presented as mean ± SD ( n = 3). Statistical significance was determined by one-way ANOVA. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: ELISA kits for human IFN- γ (CSB-E08636h), granzyme B (CSB-E08718h), and perforin (CSB-E09313h) were acquired from Cusabio (Wuhan, China).

Techniques: Lactate Dehydrogenase Assay, Co-Culture Assay, Flow Cytometry, Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay