Fig. 7.

USP11 promotes LPA1-CD14 interaction and LPS-induced lung inflammation. a. MLE12 cells were transfected with USP11-HA plasmid for 48 h, and then cells were treated with LPA (5 μM) for 30 min. Cell lysates were subjected to immunoprecipitation with LPA1 antibody, followed by CD14 and LPA1 immunoblotting. Input lysates were analyzed by immunoblotting with CD14, HA, LPA1, and β-actin antibodies. b. MLE12 cells were transfected with usp11 shRNA for 72 h, and then cells were treated with LPS (10 μg/ml) for 30 min. Cell lysates were analyzed by immunoblotting with p-Erk1&2, Erk1&2, and USP11 antibodies. Representative immunoblots were from at least three independent times. c. MLE12 cells were transfected with Usp11 shRNA for 72 h, and then cells were treated with LPS (10 μg/ml) for 3 h. Cell culture supernatants were collected and KC levels in the media were analyzed by Elisa. d–h. C57/BL6 mice (6–8/group) were intratracheally infected with control lentivirus (Lenti cont, 1 × 10⁹ cfu/mice) or lentiviral vector carrying usp11 shRNA (Lenti USP11 shRNA Lenti cont, 1 × 10⁹ cfu/mice) for 7 days, followed by i.t. administration with LPS (2 mg/kg body weight) for 24 h. Lung tissue lysates were analyzed by immunoblotting with USP11 and β-actin antibodies (d). Lung tissue were fixed and stained with H&E (e). BAL was collected, and then protein levels were examined by protein assay (f), IL-6 (g), and KC (h) were examined by Elisa.