FIGURE 5.

The effect of TLR4 on the role of E2 in Neisseria meningitidis-infected cells. (A–D) TLR4 shRNA was transfected in to HBMECs (A) and MECs (C) infected with N. meningitidis. The cells infected with N. meningitidis without TLR4 shRNA transfection and the cells transfected with negative control were used as controls. Proteins were extracted and the level of TLR4 was detected using western blotting. β-actin was used as the internal reference. The changes in protein levels in HBMECs (B) and MECs (D) were analyzed using Image-J software, and the gray values were analyzed statistically using GraphPad Prism software. (E,F) A TLR4 shRNA was transfected into E2-N. meningitidis co-stimulated HBMECs (E) and MECs (F). The E2-N. meningitidis co-stimulation experimental group without shRNA transfection and the experimental group only infected with N. meningitidis were used as controls. The supernatants of the cells were collected for ELISA assays, and the levels of IL-6, IL-8, and TNF-α in the supernatants were detected. (G–J) HBMECs (G) and MECs (I) were treated as shown. The expression of TNF-α was detected using western blotting. β-actin was the internal reference. The changes in protein levels in both cells (H,J) were analyzed as above. *P < 0.05, **P < 0.01, ***P < 0.001.