Figure 3.

CBNPs induce acute necrosis of alveolar macrophages in vitro. (A-B) The detection of CBNP-induced necrotic cells in vitro by flow cytometry with Annexin V and PI staining. (C) Primary alveolar macrophages were treated with CBNPs (100 µg/cm²) for 30 min and then stained with PI. Scale bar = 10 µm. (D) Primary peritoneal macrophages were treated with CBNPs (100 µg/cm²) for 30 min and then stained with PI. Scale bar = 10 µm. (E-F) MH-S cells were treated with CBNPs at 100 µg/cm² for 2 h. (E) Lactate dehydrogenase (LDH) release in culture supernatant was detected. (F) Morphological changes and trypan blue staining. Scale bar = 20 µm. (G) The percentages of necrotic cells after treatment with CBNPs at 100 µg/cm² for 24 h detected by Annexin V/PI. (H) Peritoneal macrophages were treated with CBNPs at 100 µg/cm² for 24 h and then stained with PI. Scale bar = 40 µm. Data are mean ± SEM; ns: no significance, ***P < 0.001 compared with control group by Student's t-test.