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Depletion of USP30 in ECs ameliorates endotoxin‐induced acute lung injury. A) Scheme shows the targeting strategy for generating the EC‐USP30KO mice. B) Immunoblotting analysis of USP30 from isolated lung ECs from Wt and EC‐USP30KO mice. C) USP30 expression was deleted in CD31‐positive cells in EC‐USP30KO mouse lungs, compared to wild‐type (Wt) mice. IgG was used as a negative control. Br, Bronchus. D) Isolated lung ECs (from Wt and EC‐USP30KO mice) were cultured on ECIS chambers to confluence. TEER was measured for cell permeability. Data shown as mean ± SEM, n = 3. E–H) Mice were challenged with (i.t.) LPS (5 mg kg −1 body weight) for 24 h. Protein levels (E), IL‐6 (F), IL‐1β (G), and neutrophil influx (H) in BAL were measured. Data shown as mean ± SEM, n = 6. I) H&E staining of lung tissues. Scale bar, 50 µm. J) Evans Blue in vivo leakage assay after (i.t.) LPS. K) Lung wet/dry ratio was measured. L) Mice were challenged with (i.p.) LPS (2 mg kg −1 body weight) for 24 h. Evans Blue in vivo leakage assay after (i.p.) LPS. Data shown as mean ± SEM, n = 6. M,N) IL‐1β and IL‐6 in BAL were measured by ELISA. Data shown as mean ± SEM, n = 6. O,P) IL‐1β and IL‐6 in plasma were measured by ELISA. Data shown as mean ± SEM, n = 6.

Journal: Advanced Science

Article Title: Activation of USP30 Disrupts Endothelial Cell Function and Aggravates Acute Lung Injury Through Regulating the S‐Adenosylmethionine Cycle

doi: 10.1002/advs.202512807

Figure Lengend Snippet: Depletion of USP30 in ECs ameliorates endotoxin‐induced acute lung injury. A) Scheme shows the targeting strategy for generating the EC‐USP30KO mice. B) Immunoblotting analysis of USP30 from isolated lung ECs from Wt and EC‐USP30KO mice. C) USP30 expression was deleted in CD31‐positive cells in EC‐USP30KO mouse lungs, compared to wild‐type (Wt) mice. IgG was used as a negative control. Br, Bronchus. D) Isolated lung ECs (from Wt and EC‐USP30KO mice) were cultured on ECIS chambers to confluence. TEER was measured for cell permeability. Data shown as mean ± SEM, n = 3. E–H) Mice were challenged with (i.t.) LPS (5 mg kg −1 body weight) for 24 h. Protein levels (E), IL‐6 (F), IL‐1β (G), and neutrophil influx (H) in BAL were measured. Data shown as mean ± SEM, n = 6. I) H&E staining of lung tissues. Scale bar, 50 µm. J) Evans Blue in vivo leakage assay after (i.t.) LPS. K) Lung wet/dry ratio was measured. L) Mice were challenged with (i.p.) LPS (2 mg kg −1 body weight) for 24 h. Evans Blue in vivo leakage assay after (i.p.) LPS. Data shown as mean ± SEM, n = 6. M,N) IL‐1β and IL‐6 in BAL were measured by ELISA. Data shown as mean ± SEM, n = 6. O,P) IL‐1β and IL‐6 in plasma were measured by ELISA. Data shown as mean ± SEM, n = 6.

Article Snippet: USP30 , Mouse / IgG1 , 1:1000 , Santa Cruz, sc‐515235.

Techniques: Western Blot, Isolation, Expressing, Negative Control, Cell Culture, Permeability, Staining, In Vivo, Enzyme-linked Immunosorbent Assay, Clinical Proteomics