Extended Data Fig. 7: MICL recognizes NETs.
From: Recognition and control of neutrophil extracellular trap formation by MICL
a, NET-bound Sytox green fluorescence of bone marrow-isolated neutrophils stimulated with preformed murine NETs (mNETs). Data is a representative example of n = 3 independent experiments, mean ± SD performed in triplicate. b, Mean fluorescence intensity (MFI) of cit-H3 and MPO (mean ± SD, 3 fields of view per condition) during NET formation in WT or MICL-deficient mNETs-stimulated neutrophils. Statistical significance determined by Student’s t-test. c, ROS production of MICL-deficient neutrophils mNETs-stimulated with or without polyxymin B. Data is a representative example of n = 2 independent experiments, mean ± SD performed in triplicate. d, NET formation and quantification of area under curve (AUC, right) of WT or MICL-deficient mNETs-stimulated neutrophils with GSK484 or DPI. Data is a representative example of n = 2 independent experiments, mean ± SD performed in triplicate. e, Fold change in MFI of MICL expression on cultured neutrophils (CD66b+CD15+), relative to human neutrophils treated with negative control scrambled guide RNA. f, ROS production by preformed NETs of human MICL-knockout cultured neutrophils (hKO) or human WT derived from CD34+ haematopoietic progenitors. Data is a representative example of n = 2 independent experiments, mean ± SD performed in triplicate. g, Fc-MICL NET recognition by ELISA in the presence/absence of anti-MICL antibodies. Pooled data represented as mean ± SD of n = 2 independent experiments performed in triplicate. Statistical significance determined by One-way ANOVA and Bonferroni multiple comparison test. h, ROS production of MICL-deficient neutrophils stimulated with untreated NETs, Proteinase K-treated NETs or DNaseI-treated NETs. Data is a representative example of n = 3 independent experiments, mean ± SD performed in triplicate. i, Fc-MICL genomic DNA recognition by ELISA in the presence/absence of anti-MICL antibodies. Pooled data represented as mean ± SD of n = 2 independent experiments performed in triplicate. Statistical significance determined by One-way ANOVA and Bonferroni multiple comparison test. j, Cell free DNA concentration 4 h after NETs or LPS intraperioteneal injection in WT and MICL-deficient animals (n = 1 experiment with 6 animals/group). *, p < 0.05.
