(A) The HMGB1 release in the supernatant medium in MLE-12 (upper) and organoids (lower) treated with 8% CSE in combination with 10 μM Z-VAD (Z) or 5 μM GSK’872 (G) for 16 hours. (B and C) MLE-12 cells were treated as in A and stained with CellTrace Far Red (red), then cocultured with BMDMs stained with CellTrace CFSE (green) for 8 hours. BMDM phagocytosis was analyzed by flow cytometry (B) and confocal microscopy (C). Representative phagocytosis was shown. Original magnification, ×1000. (D and E) MLE-12 cells treated as in B were cocultured with BMDMs. The mRNA level of TNF-α (D) and IL-6 (E) in MLE-12 cells or MLE-12 and BMDM coculture system was analyzed. Each experiment was repeated 3 times. Data represent the means ± SEM. *, P < 0.05; **, P < 0.01. One-way ANOVA with Tukey’s multiple-comparison test was conducted. N/A, untreated BMDMs.