Fig. 2: epiCaspase-1 suppresses immunogenic cell death and promotes granulocyte-monocyte progenitors distally in the bone marrow.

a Representative of three western blot replicates confirming the efficiency of caspase-1 knockout by CRISPR-Cas9 in G69 bladder cancer cells and knockdown by shRNA in 4T1 breast cancer cells. b LDH release from wild-type (WT) and caspase-1 knockout (Casp1KO) G69 cells after gemcitabine treatment (3 biological replicates). c LDH release from wild-type (WT) and caspase-1 knockdown (shCasp1) 4T1 cells after gemcitabine treatment (3 biological replicates). d Schematic diagram illustrating the gold-standard vaccination assay for evaluating immunogenic cell death in vivo. Created in BioRender. Wong, S. (2026) https://BioRender.com/az159ga. e Kaplan-Meier plot showing tumor-free incidence of mice vaccinated with either vehicle, gemcitabine treated G69 wild-type (gemG69^WT), or gemcitabine treated caspase-1 KO (gemG69^Casp1KO) cancer cells as “vaccines” (n = 10 mice per group). f Corresponding changes in tumor burden from (e). g Kaplan-Meier plot showing tumor-free incidence of mice vaccinated with either vehicle (n = 10 mice per group), gemcitabine treated 4T1 wild-type (gem4T1^WT, n = 10), or gemcitabine treated caspase-1 knockdown (gem4T1^shCasp1, n = 8) cancer cells as “vaccines”. h Corresponding changes in tumor burden from (g). i Kaplan-Meier plot showing tumor-free incidence of mice vaccinated with either vehicle, gemG69^WT, or gemG69^Casp1KO cancer cell “vaccines” with prior treatment anti-CD8 mAb (αCD8) (n = 5 mice per group). j Kaplan-Meier plot showing tumor-free incidence of mice vaccinated with gemG69^WT cancer cell as “vaccines” in mice depleted (αLy6G) and not depleted of neutrophils (n = 8 mice per group). k Schematic diagram depicting a modified vaccination assay in vivo. Red arrow: vaccination doses and treatment scheme. Created in BioRender. Wong, S. (2026) https://BioRender.com/6n2awul. l Flow cytometry gating strategy for the analysis of bone marrow progenitor cell subpopulations, following vaccination with vehicle [black], gemG69^WT [red] or gemG69^Casp1KO [blue] as “vaccines”. m Flow cytometry results showing the counts of granulocyte-monocyte progenitors (GMPs; Lin^-Sca^-cKit⁺CD16/32^hiCD34^hi) in mice vaccinated with gemG69^WT, or gemG69^Casp1KO (n = 6 mice per group). Statistics: Ordinary one-way ANOVA followed by Dunnett’s multiple comparison test (b, c, m), two-way ANOVA with Geisser-Greenhouse correction followed by Dunnett’s multiple comparison test (f, h), and log-rank (Mantel-Cox) test (e, g, i, j) were performed. Error bars represent standard error of mean (s.e.m.) unless otherwise stated. Source data are provided as a Source Data file.