Macrophages (RAW264.7) were performed to confirm the effect of SA primed E. coli on inflammatory responses. Macrophages were cultured in 1640 medium (Sigma-Aldrich, MO, USA) suppled with 10% fetal bovine serum (BI, Israel) and 1% ampicillin and streptomycin at 37 ℃ with 5% CO2. For the E. coli stimulation experiment, cells (106 cells/mL) were incubated in 24-well plates and removed the ampicillin and streptomycin (Sigma Aldrich, MO, USA), and then cells were treated with Neu5Ac or glucose-cultured E. coli (106, 107, and 108 CFU) for 24 h. Finally, the supernatants were harvested and proinflammatory cytokines were determined by ELISA.
Zhao C., Hu X., Qiu M., Bao L., Wu K., Meng X., Zhao Y., Feng L., Duan S., He Y., Zhang N, & Fu Y. (2023). Sialic acid exacerbates gut dysbiosis-associated mastitis through the microbiota-gut-mammary axis by fueling gut microbiota disruption. Microbiome, 11, 78.
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