Fig. 1

MPO expression and HOCl production in PC12 cells under monoculture or co-cultured with microglial BV2 cells after exposure to normoxic conditions (NC) or OGD/R in vitro. PC12 cells were mono-cultured or transwell co-cultured with BV2 cells and then exposed to 4 h OGD (glucose-free medium with 1% O2/5% CO2) or normal conditions (NC, normal glucose medium with 21% O2/5% CO2), and then the cells were cultured with fresh normal glucose medium with 21% O2/5% CO2 and incubated for 24 h. The cells were treated with 4-ABAH (50 µM) at the onset of reoxygenation. A, B Western blot analysis for MPO expression in PC12 cells under monoculture or co-culture with BV2 cells conditions. C, D HKOCl-3 staining HOCl production from PC12 cells in single cultured plates or co-cultured transwells with BV2 cells, bar = 200 μm. E Quantification of MPO level in the supernatant by ELISA. F Necrotic cell death was detected by LDH release from PC12 cells. G, H Annexin V-FITC/PI staining flow cytometry assay for apoptotic cell death. Transwell-OGD/R: BV2 and PC12 co-cultured cells were exposed to 4 h OGD with 24 h of reoxygenation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ##p < 0.01, ####p < 0.0001. All data are presented as mean ± SEM. (Statistical methods: B, C, E, F one-way ANOVA followed by Tukey’s multiple comparisons test. G Two-way ANOVA followed by Bonferroni multiple comparisons test.)