Fig. 6: PRL2 related NET formation contributes to the pathology in iRBCs induced ALI.

a A schematic showing the experimental design for iRBCs and TNF-α induced acute lung injury (ALI) model. b–g C57BL/6 J mice were induced ALI in (a) or grouped as normal control (Con) (n = 6 mice per group). b Representative images of lung tissues stained with hematoxylin-eosin (H&E). Scale bars, left: 50 μm, right: 10 μm. c Pulmonary pathology scores and infiltrated neutrophil numbers were quantified from (b). d Quantification of neutrophils and neutrophil extracellular traps (NETs) in immunofluorescence images of lungs. e Representative immunofluorescence images of lungs as described in (d). DNA is stained in blue (DAPI), myeloperoxidase is stained in green (MPO) and citrullinated histone H3 is stained in red (Cit-H3). Scale bars, 20 μm. Neutrophils are indicated as co-stained with MPO and DAPI. NETs are indicated as co-stained with MPO and Cit-H3. f Percentage of different subsets of peripheral blood cells. g Relative PRL2 mean fluorescence intensity (MFI) in different subsets of peripheral blood cells, normalized to normal mice. h A schematic showing the experimental design for neutrophil depletion in iRBCs and TNF-α induced ALI model. i–m Wildtype (WT) and PRL2 myeloid cell conditional knockout (CKO) mice were intraperitoneally injected with anti-Ly6G monoclonal antibody or an isotype control 24 h before iRBCs injection (n = 6 mice per group). i Number of peripheral neutrophils from the four groups of mice 24 h after iRBCs injection. j Representative images of lung tissues stained with H&E. Scale bars, left: 50 μm, right: 10 μm. k Pulmonary pathology scores and infiltrated neutrophils numbers were quantified from (j). l Quantification of neutrophils and NETs in immunofluorescence images of lung tissues. m Representative immunofluorescence images of lung tissues. Staining panel is as described in (e). All data were pooled from two independent experiments. Data are presented as the mean ± SEM or in violin plots showing the median and interquartile range. p values were calculated by two-tailed unpaired t test (c up, f, g), two-tailed Mann–Whitney test (c down, d) or two-way ANOVA with Tukey’s multiple testing (i, k, l) and shown in the figures. Source data are provided as a Source Data file.