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. 2019 Feb 6;5(2):eaav2309. doi: 10.1126/sciadv.aav2309

Fig. 5. 4-PBA enhances peroxisome homeostasis in neutrophils.

Fig. 5

Neutrophils were purified from the BM of wild-type C57BL/6 mice and treated with PBS, superlow-dose LPS (100 pg/ml), and/or 4-PBA (1 mM) for 2 days. (A) Representative histogram of ROS level determined by CellROX labeling. (B) Quantification of ROS levels in neutrophils (n = 3 for each group). (C) Representative confocal microscopy images of the neutrophils stained with anti-PMP70 (peroxisomal membrane protein 70) and anti-LAMP1 (lysosomal-associated membrane protein 1) antibodies to demonstrate the localization and fusion of peroxisomes and lysosomes. Scale bars, 5 μm. (D) Western blot data of oxCaMKII and 5-LOX expression. (E) Representative histograms and quantification of p-STAT1 (n = 3 for each group), ATF4, and KLF2 (n = 5 for each group) levels as determined by flow cytometry. (F) Surface phenotype of neutrophils was analyzed by flow cytometry (n = 3 for each group). (G) The levels of MPO, MMP9, and LTB4 were determined by ELISA (n = 3 for each group). Data are representative of three independent experiments, and error bars represent means ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, one-way ANOVA.