Starch broth

The murine macrophage cell line Raw264.7 (used within 10 passages) was purchased from American Type Culture Collection (ATCC) and cultured in Dulbecco’s modified Eagle’s medium (Gibco, Life Technologies, Germany) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin-streptomycin (Solarbio, Beijing, China) at 37 °C in a 5% CO2 humidified chamber. Human pulmonary microvascular endothelial cells (HPMECs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) and cultured in the specialized medium (ScienCell, USA). Raw264.7 cells and HPMECs were pretreated with BP-1-102 (5 μM, dissolved in DMSO) for 2 h and then stimulated with LPS (1 μg/ml) for 0.5 or 6 h.
Peritoneal macrophages (PMs) were isolated from 6- to 7-week-old C57BL/10ScNJ and C57BL/6 mice as previously described [32 ]. Briefly, mice were intraperitoneally injected with 2 ml of starch broth (Sigma, USA), and then the peritoneal cavity was washed with RPMI medium (Gibco, Life Technologies, Germany) three times to isolate peritoneal exudate cells. After 2 h of culture, the adhered cells were used as macrophages. PMs were stimulated with LPS (1 μg/ml) for 6 h for further experiments.

Human pulmonary microvascular endothelial cells (HPMECs, 3000, ScienCell, USA) were cultured in endothelial cell medium (ECM, 1001, ScienCell, USA), matched with 5% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified incubator under 5% CO₂ at 37 °C. The HPMECs were seeded into the culture dish for one night. The medium was replaced with fetal bovine serum (FBS) medium for 6–8 h. The HPMECs were cultured with 1 μg/ml LPS with or without 50 ng/ml of FGF2 for 0.5 h or 6 h.
Peritoneal macrophages (PMs) were isolated from 6–8 weeks old male C57BL/6 mice. Briefly, mice were intraperitoneal injected with 2 ml Starch broth (Sigma, USA), and then cold RPMI 1640 medium was used to wash the peritoneal cavity several times to isolate peritoneal exudate cells. The cells were cultured in a humidified incubator for 2 h and the adhered cells were collected as macrophages for further experiments. PMs were cultured with 1 μg/ml LPS with or without 50 ng/ml of FGF2 for 0.5 h or 6 h.