Fig. 7: The programmed death ligand-1(PD-L1)/cluster of differentiation 80 (CD80)/mitogen-activated protein kinase (MAPK) axis mediates Tudor domain-containing protein 9 (TDRD9)-dependent cuproptosis regulation.

A Neutrophils were pre-transducted with lentiviral-based short hairpin RNA (shRNA) knocking down TDRD9 (sh-TDRD9) and lentiviral PD-L1 overexpression construct (oe-PD-L1), followed by treatment with 10 μM elesclomol (ES) for 24 h and Pseudomonas aeruginosa (PA) strain PAO1 at a multiplicity of infection (MOI) of 10 for 1 h. Quantitative analysis of colony-forming unit (CFU) in neutrophils. n = 3 independent experiments. B PD-L1 overexpression rescued TDRD9 knockdown-induced pathway suppression. n = 3 independent experiments. C Enzyme-linked immunosorbent assay (ELISA) analysis of PD-L1 and sCD80 levels in cell culture medium. n = 3 independent experiments. D Viability restoration by PD-L1 overexpression in TDRD9-depleted neutrophils. n = 3 independent experiments. E Cuproptosis-related protein dynamics under TDRD9 knockdown ± PD-L1 overexpression. n = 3 independent experiments. F ELISA analysis of tumor protein p53 (p53) levels in cell culture medium. n = 3 independent experiments. G Neutrophils were pre-transducted with oe-PD-L1 and lentiviral-based shRNA knocking down CD80 (sh-CD80), followed by treatment with 10 μM elesclomol (ES) for 24 h and infection with PAO1 at an MOI of 10 for 1 h. Signaling pathway-related protein expressions under PD-L1 overexpression ± CD80 knockdown. n = 3 independent experiments. H ELISA analysis of PD-L1 and sCD80 levels in cell culture medium. n = 3 independent experiments. I Viability reduction by CD80 knockdown in PD-L1-overexpressed neutrophils. n = 3 independent experiments. J, K Cuproptosis-related protein dynamics under PD-L1 overexpression ± CD80 knockdown. n = 3 independent experiments. L PD-L1-CD80 interaction using co-immunoprecipitation assay. n = 3 independent experiments. Data represent median ± interquartile range (IQR), and were analyzed by two-sided one-way ANOVA with Tukey’s multiple comparisons test (A–K). Source data are provided as a Source Data file.