[Retracted] HD-13 Induces Swine Pneumonia Progression via Activation of TLR9

Figure 3

HD-13 activates the MAPK and NF-κB pathway. 3D4/21 cells were infected with HD-13 for 0 h, 0.5 h, 1 h, and 1.5 h, respectively. (a) Western blot was performed to assess the levels of MAPK pathway-related proteins, including ERK, p-ERK, p38, p-p38, JNK, and p-JNK. (b) The levels of NF-κB pathway-related proteins, including p65, p-p65, IKKα, and p-IKKα, were evaluated by Western blot, following infection with HD-13 for 0 h, 1 h, 6 h, and 112 h, respectively. (c) Immunofluorescence was conducted to assess p-p65 expression. 3D4/21 cells were randomly divided into 3 groups: control group—cells were untreated; HD-13 group—cells were infected with HD-13 for 1.5 h; and HD-13+PD98059/HD-13+SB203580/HD-13+SP600125/HD-13+BAY11-7082 group—cells were infected with HD-13 and 10 μM PD98059 or 10 μM SB203580 or 10 μM SP600125 or 10 μM BAY11-7082 for 1.5 h. PD98059 serves as the p-ERK/ERK pathway inhibitor, SB203580 serves as the p-p38/p38 pathway inhibitor, SP600125 serves as the p-JNK/JNK pathway inhibitor, and BAY11-7082 serves as the NF-κB pathway inhibitor. (d, e) RT-qPCR was performed to assess the mRNA levels of IL-1β, IL-6, IL-12, and TNF-α. (f, g) ELISA was carried out to detect the expression of IL-1β, IL-6, IL-12, and TNF-α in the supernatant of 3D4/21 cells. , , and vs. control group; ^#, ^##vs. HD-13 group.

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