Fig. 2: PD-L1 governs the mobilization of neutrophils through regulating their autocrine secretion of CXCL1/2.

a Number of up-regulated genes in wild-type mu-PMNs and down-regulated genes in Clec7a^−/− and Cd274^−/− mu-PMNs stimulated with curdlan. b GO analysis of 38 down-regulated genes in Clec7a^−/− and Cd274^−/− mu-PMNs stimulated with curdlan. c Heatmaps show of down-regulated genes involved in positive regulation of cell migration as shown in b and in Hu-PMNs treated with anti-PD-L1(αPD-L1, 10 μg/ml), which were then stimulated with curdlan for 4 h. d, e ELISA quantification of CXCL1 and CXCL2 in the supernatant of Clec7a^−/− or Cd274^−/− mu-PMNs, which were stimulated with curdlan (25 μg/well) for 4 h. f Immunofluorescence staining of nuclear PD-L1 location in mu-PMNs stimulated with curdlan (25 μg/well) for 4 h and Hu-PMNs stimulated with curdlan (50 μg/well) for 6 h. Scale bar = 5 μm. g Representative histological images with immunoblotting assay of PD-L1 in mu-PMNs and Hu-PMNs, which were stimulated with curdlan for 6 h. h ELISA quantification of CXCL1 and CXCL2 in the supernatant of mu-PMNs, which were stimulated with curdlan (25 μg/well) and ivermectin (20 μM) for 4 h. i ELISA quantification of CXCL1 and CXCL2 in the supernatant of wild-type or Cd274^−/− mu-PMNs, which were stimulated with curdlan (25 μg/well) and inhibitor Plicamycin (5 nM) for 4 h. j Individual GSEA plots of neutrophil migration pathway in two independent RNA-Seq data of wild-type and Cd274^−/− mu-PMNs, which were stimulated with curdlan. P values were calculated by hypergeometric test and adjusted for multiple comparisons. k Schematic diagram of trans-well migration assay. The lower chamber contains the supernatant of neutrophils in different groups, which were stimulated with curdlan (25 μg/well for mu-PMNs and 50 μg/well for Hu-PMNs) for 4 h. l–o Trans-well migration assay of naive neutrophils driven by the supernatants of curdlan-stimulated wild-type, Clec7a^−/− or Cd274^−/− neutrophils combined with treatment of anti-CXCL1 (αCXCL1, 0.2 µg/ml), anti-CXCL2 (αCXCL2, 2 µg/ml), AZD-5069 (0.8 nM) or ivermectin (20 μM), and human neutrophils combined with treatment with anti-Dectin-1(αDectin-1, 1 μg/ml), anti-PD-L1(αPD-L1, 10 μg/ml). Data were presented as mean ± SD; n = 3 (d, e, h, i, l–o) biological independent samples. Data were analyzed by unpaired two-sided Student’s t test in d, e, h, l–o or one-way ANOVA adjusted for multiple comparisons in i. Source data are provided as a Source data file.