Anti-TGEV Miller Strain Infection Effect of Lactobacillus plantarum Supernatant Based on the JAK-STAT1 Signaling Pathway

Wang, Kai; Ran, Ling; Yan, Tao; Niu, Zheng; Kan, Zifei; Zhang, Yiling; Yang, Yang; Xie, Luyi; Huang, Shilei; Yu, Qiuhan; Wu, Di; Song, Zhenhui

Lp-1s was prepared as above and then diluted to an OD 600 of 0.45, 0.225, 0.113, and 0.056 with RPMI 1640 high-sugar medium. IPEC-J2 cells were seeded at 1.8 × 10 6 /mL in 6-well plates and cultured in 5% CO 2 at 37 • C. At 90% confluence, the medium was discarded, the cells were washed three times with PBS, 1 mL of each gradient dilution of Lp-1s was added to each well, and the cells incubated at 37 • C in 5% CO 2 for 90 min. The medium in the control group was replaced with RPMI 1640. For the other wells, the supernatant was discarded, TGEV (multiplicity of infection (MOI) = 0.1) in RPMI 1640 medium was added to Lp-1s-treated IPEC-J2, and incubated at 37 • C in 5% CO 2 for 1.5 h. The supernatants were discarded and incubation continued in RPMI 1640 high glucose medium. After 48 h of culture, proteins were extracted for western blotting to detect the levels of the TGEV N protein after treatment with different concentrations of Lp-1s.