Journal: bioRxiv

Article Title: Inhibition of 3-mercaptopyruvate sulfurtransferase enhances CD8⁺ T-cell antitumor immunity

doi: 10.1101/2025.09.22.675323

Figure Lengend Snippet: Renca-H2-Kb GFP⁺ cells were treated with increasing concentrations of HMPSNE. (A) Live-cell imaging expressed as GFP signal normalized to the starting point (T₀ = 1) for each well. Data represent mean ± S.E.M. from one representative experiment out of three with three technical replicates. (B) Metabolic activity measured by CellTiter-Glo® assay at 24 h and 48 h. (C) Representative images of Renca-H2-Kb GFP⁺ cells treated with increasing concentrations of HMPSNE for 72 h. Images were acquired using the Incucyte® Live-Cell Analysis System (20× objective). (D) Cell proliferation determined by BrdU incorporation. (E) Annexin V-positive, PI-negative apoptotic cells measured by flow cytometry. (F) Expression of CD70, CD86, and PD-L1 (mean fluorescence intensity, MFI) on Renca-H2-Kb GFP⁺ cells determined by flow cytometry. Data represent mean ± SD from three independent experiments. Statistical analysis was performed using two-way ANOVA with Tukey’s post hoc test for (A) and Dunnett’s post hoc test for (B, D-E). ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05 versus vehicle control (0 µM HMPSNE).

Article Snippet: Anti-PD-L1 antibody (10 μg/mL; InvivoGen, #pdl1-mab15-1) was added where indicated.

Techniques: Live Cell Imaging, Activity Assay, Glo Assay, Cell Analysis, BrdU Incorporation Assay, Flow Cytometry, Expressing, Fluorescence, Control

Journal: bioRxiv

Article Title: Inhibition of 3-mercaptopyruvate sulfurtransferase enhances CD8⁺ T-cell antitumor immunity

doi: 10.1101/2025.09.22.675323

Figure Lengend Snippet: CD8⁺ T cell-mediated cytotoxicity was assessed using Renca-H2-Kb GFP⁺ tumor cells either loaded with SIINFEKL or left unloaded, and co-cultured with CD8⁺ T cells isolated from OT-1 mice in the presence of increasing concentrations of HMPSNE. (A) Proliferation curves of tumor cells based on GFP signal, normalized to the starting point (T₀ = 1) for each well. (B) Tumor cell proliferation over 114 h with increasing concentrations of HMPSNE and/or anti-PD-L1, expressed as values normalized to T₀. Data represent mean ± SEM from one representative experiment out of three with three technical replicates. Statistical analysis was performed using two-way ANOVA with Tukey’s post hoc test. ****p < 0.0001; ***p < 0.001; *p < 0.05 versus indicated sample.

Article Snippet: Anti-PD-L1 antibody (10 μg/mL; InvivoGen, #pdl1-mab15-1) was added where indicated.

Techniques: Cell Culture, Isolation