Journal: Nature Communications
Article Title: Plasma redox imbalance caused by albumin oxidation promotes lung-predominant NETosis and pulmonary cancer metastasis
Figure Lengend Snippet: Plasma redox imbalance induces NETosis. a Concentration of free thiol in plasma from wild-type (WT) vs. albumin deficient (Alb −/− ) C57BL/6 mice and control vs. albumin-depleted plasma from NOD scid gamma (NSG) and BALB/c mice. b – d Plasma isolated from the blood of WT and Alb −/− C57BL/6 mice and NSG mice 2 days post saline, IAA (30 mg/kg), or LPS (8 mg/kg) injection was subjected to FPLC ( b , c ) and Western blotting ( d ). b Representative FPLC results for the redox state of albumin in C57BL/6 (left) and NSG (right) mice. c The ratio of peak intensity for reduced/oxidized albumin in NSG mice. d Western blotting with an antibody against C-reactive protein (CRP) and serum amyloid P component (SAP). e – g Plasma was isolated from the blood of WT and Alb −/− C57BL/6 mice and NSG mice 2 days post saline or IAA (30 mg/kg) injection following intravenous injection of PBS or murine albumin (20 mg/mouse). e , f Immunoprecipitation of cell-free DNA in the plasma from C57BL/6 ( e ) and NSG ( f ) mice using an anti-dsDNA antibody followed by Western blotting using an antibody against citrullinated histone H3 (Cit-H3). g Plasma free thiol concentration. a , c , g Results represent individual values with the mean ± s.d. ( n = 3–6; biological replicates, * P
Article Snippet: We used LL-37 (1:1,000, OSC00009W, Osenses Pty Ltd., Australia), histone H3 (citrulline R2 + R8 + R17, 1:500, ab5103, Abcam, MA, USA), transferrin (1:500, ab82411, Abcam, MA, USA), β-actin (1:200, sc-47778 HRP, Santa Cruz, CA, USA) where indicated.
Techniques: Concentration Assay, Mouse Assay, Isolation, Injection, Fast Protein Liquid Chromatography, Western Blot, Immunoprecipitation