Primary antibody against nrf2

HPMECs were seeded into the culture dish with coverslips at a density of 1×10⁵ cells/mL. After 24 h, HPMECs were fixed with 4% paraformaldehyde for 15 min and permeabilized with 0.5% Triton X-100 for 20 min at room temperature. The dishes were then washed with phosphate-buffered saline (PBS) for three times, followed by incubation with 3% bovine serum album (BSA) for 30 min at room temperature. Primary antibody against Nrf2 (Proteintech; Wuhan, China) (1:100 dilution in PBS with 1% BSA) was then added overnight at 4°C. Cells were washed with cold PBS for three times and then incubated in Alexa 488-conjugated secondary antibody. The nuclei were stained with DAPI solution (1 mg/mL) for 5 min at room temperature. The slides were washed again and the images were collected on a zeiss LSM880 confocal microscope (Carl Zeiss, German).

LPS extracted from the membrane of Escherichia coli 0111:B4 was purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium propionate (SP) (HPLC≥99%) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Serum and lung superoxide dismutase (SOD) and hematoxylin and eosin were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). For immunohistochemical analysis, the antibody against von Willebrand factor (vWF)  was obtained from Proteintech (Wuhan, China). For immunofluorescence staining, a primary antibody against Nrf2 was obtained from Proteintech (Wuhan, China). For Western blotting analysis, rabbit monoclonal antibody against Nrf2 was purchased from Cell Signaling Technology (Beverly, MA, USA). Rabbit polyclonal antibodies against vWF and Keap-1 were obtained from Proteintech (Wuhan, China). Mouse monoclonal antibody against GAPDH was obtained from Thermo Scientific (Waltham, MA, USA).