Journal: Cell Death and Differentiation
Article Title: ARID1A mutations protect follicular lymphoma from FAS-dependent immune surveillance by reducing RUNX3/ETS1-driven FAS-expression
doi: 10.1038/s41418-025-01445-3
Figure Lengend Snippet: A Schematic of the FASLG-induced apoptosis assay (soluble human recombinant FAS ligand treatment). B Percent AnnexinV-positive OCI-Ly8 (left panel) and OCI-Ly1 (right panel) clones with ARID1A WT, het, KO and overexpression of RUNX3 on het (het + RUNX3) after 24 h treatment with increasing dose of purified soluble human recombinant FAS ligand ( N = 3). P-values are from a linear regression on square root transformed values tested against WT and Bonferroni-adjusted . C Schematic of the T-cell mediated killing assay. D Conjugate formation of OCI-Ly8 cells (CFSE + ) and CD8 + T-cells (VPD + ); double-positive conjugates (CSFE + /VPD + ) quantified as percentage of total CD8 + T-cells (VPD + ) ( N = 5) at indicated conditions. E T cell-mediated cytotoxicity assessed by quantifying the fraction of OCI-Ly8 cells (CFSE + ) undergoing apoptosis upon co-culture with CD8+ T cells, expressed as a percentage of all measured cells under the indicated conditions (N = 5, biological replicates performed with different healthy T cell donors). P-value is from the paired Welch test. Het + RUNX3 was tested against het. Pooled data from biological replicates (N) are represented as mean ± SD . F Schematic representation of the CAR T-cell mediated killing assay. G Luciferized OCI-Ly8 ARID1A WT cells were co-cultured with CD19-CAR T cells at a 1:3 effector-to-target (E:T) ratio with or without a FAS-blocking antibody or an IgG isotype control. Luminescence was measured after 24 h and normalized to the baseline luminescence at 1 h. P-values were calculated using the paired Welch test, comparing WT against WT + FAS and WT + EV. Pooled data from biological replicates (N = 3; performed with CAR T-cells from different healthy donors) are represented as mean ± SD . H Luciferized OCI-Ly8 cells of indicated genotypes were co-cultured with CD19-CAR T cells at a 1:3 effector-to-target (E:T) ratio for 48 h. ( N = 3, biological replicates performed with CAR T-cells from different healthy donors). P-values are from the paired Welch test. WT was tested against het, KO and het + ARID1A. Pooled data from biological replicates are represented as mean ± SD .
Article Snippet: Lymphoma cells were stained with 1 μM CellTrace TM CFSE Cell Proliferation Dye (Thermo Fisher Scientific, Waltham, Massachusetts, USA) for 4 min at room temperature (RT).
Techniques: Apoptosis Assay, Recombinant, Clone Assay, Over Expression, Purification, Transformation Assay, Co-Culture Assay, Cell Culture, Blocking Assay, Control
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