Fig. 3: Early and late neutrophil depletion result in opposite neurodevelopmental outcome.

A Schematic experimental design showing analysis timeline and readouts to assess the impact of neutrophil depletion on HI-induced long-term neurodevelopmental deficits. Neutrophils were depleted by combing anti-Ly6G with anti-rat IgG treatment at 12 h/36 h/50 h (early depletion) or 6/7/8 days (late depletion) after HI. B Body weight development over the time course of the experiment, two-way-repeated measurement ANOVA followed by Tukey’s multiple comparisons test. C HI-induced hyperactivity-related behavior analyzed in the Open Field test by quantification of mean velocities and mobility. Representative images reveal movement tracks over the observation period of 5 min. D Anxiety-related behavior was evaluated in the Elevated Plus Maze by measurement of the time spent in the open arms of the maze. Representative images reveal heat maps summarizing localization of mice over the observation period of 5 min. E Cresyl staining and quantification of total hemisphere atrophy 6 weeks after HI Scale bar: 1 mm. F Correlation of normalized behavioral parameters (lowest value: 0, highest value: 100) from (C) and (D) with tissue atrophy from (E), simple linear regression, linear regression lines with 95% confidential intervals are shown for each readout. G qPCR of different neuronal (NeuN, MAP2) and myelination (CC1, CPNase, MAG, MBP) transcripts. Mean fold change values compared to sham isotype treatment are represented. Individual data and results for comparisons between experimental groups are presented in Supplementary Fig. S5E and the source data file. H Western blot analysis of MBP protein expression, normalized to the total protein. Uncropped and unprocessed images of gels and blots are provided in Supplementary Fig. S6. Bar graphs in C/D/E/H show median values, one-way ANOVA followed by Holm-Šídák’s multiple comparisons test. n = 8 sham isotype (4 early, 4 late), n = 8 sham anti-Ly6G (4 early, 3 late), n = 15 HI isotype (8 early, 7 late), n = 11 HI anti-Ly6G early, n = 13 HI anti-Ly6G late. *p < 0.05, **p < 0.01, ***p < 0.001. Individual values and exact p values are given in the source data file.