Journal: Lipids in Health and Disease

Article Title: Exosomal IL-8 derived from Lung Cancer and Colon Cancer cells induced adipocyte atrophy via NF-κB signaling pathway

doi: 10.1186/s12944-022-01755-2

Figure Lengend Snippet: Identification of EVs isolated from LLC and C26 cells. A Morphological analysis of EVs derived from LLC and C26 cells by electron microscopy (bars = 100 nm). B Evaluating the particle size of EVs based on Nanoparticle Tracking Analysis (NTA). C Detection of EVs markers and IL-8 by Western blot. D 3 T3-L1 adipocytes were co-cultured with Dil-labeled LLC-EVs and C26-EVs (red) and fixed for confocal imaging. Scale bar =50 μm. E Kaplan–Meier analyses indicating reverse correlation between IL-8 expression in tumor tissues and overall survival of lung cancer cohorts

Article Snippet: Reagents and sources: recombinant mouse CXCL15/Lungkine protein (442-LK-002, R&D Systems); anti-IL-8 (AF-208-SP, R&D Systems); CXCR2 inhibitor (SB225002, MCE); NF-κB inhibitor (BAY 11-7082, Sigma-Aldrich); Oil red O solution (Servicebio, China).

Techniques: Isolation, Derivative Assay, Electron Microscopy, Western Blot, Cell Culture, Labeling, Imaging, Expressing

Journal: Lipids in Health and Disease

Article Title: Exosomal IL-8 derived from Lung Cancer and Colon Cancer cells induced adipocyte atrophy via NF-κB signaling pathway

doi: 10.1186/s12944-022-01755-2

Figure Lengend Snippet: IL-8 in EVs induces lipolysis partly by activating NF-κB signaling pathway. A-C Western blot and the densitometric analysis of phosphorylated-p65, nuclear p65, total p65, UCP1, PGC1α in 3 T3-L1 adipocyte treated with IL-8, LLC-EVs (10 μg), C26-EVs (10 μg) and/or anti-IL-8 ( n = 3), as indicated. D The activation of p65 NF-κB (as marked in Green) in 3 T3-L1 adipocytes was confocally imaged upon co-culturing with IL-8 (with and without specific anti-IL-8), LLC-EVs and/or C26-EVs (with and without specific anti-IL-8). The nucleus (Blue) was labeled with DAPI and 10 μg EVs were applied in test. E Oil red O staining assay for above groups. Bar = 100 μm. F Relative levels of glycerol release in cell supernatants upon above treatments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Reagents and sources: recombinant mouse CXCL15/Lungkine protein (442-LK-002, R&D Systems); anti-IL-8 (AF-208-SP, R&D Systems); CXCR2 inhibitor (SB225002, MCE); NF-κB inhibitor (BAY 11-7082, Sigma-Aldrich); Oil red O solution (Servicebio, China).

Techniques: Western Blot, Activation Assay, Labeling, Staining

Journal: Lipids in Health and Disease

Article Title: Exosomal IL-8 derived from Lung Cancer and Colon Cancer cells induced adipocyte atrophy via NF-κB signaling pathway

doi: 10.1186/s12944-022-01755-2

Figure Lengend Snippet: CXCR2 inhibitors partially relieved adipocyte wasting caused by IL-8 and EVs. A The viability of 3 T3-L1 adipocytes treated with different doses of CXCR2 inhibitor SB225002 as assessed by CCK8. B-D Western blot analyses of phosphorylated-p65, total p65, UCP1, and PGC1α in adipocytes co-cultured with IL-8 (with and without SB225002), LLC-EVs and/or C26-EVs (with and without SB225002) ( n = 3), as indicated. 10 μg EVs were used in this experiment. E Oil red O staining test of the above groups. Bar =100 μm. F Relative levels of glycerol release in cell supernatants with the above treatment. 10 μg EVs were used in this experiment. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Reagents and sources: recombinant mouse CXCL15/Lungkine protein (442-LK-002, R&D Systems); anti-IL-8 (AF-208-SP, R&D Systems); CXCR2 inhibitor (SB225002, MCE); NF-κB inhibitor (BAY 11-7082, Sigma-Aldrich); Oil red O solution (Servicebio, China).

Techniques: Western Blot, Cell Culture, Staining

Journal: Lipids in Health and Disease

Article Title: Exosomal IL-8 derived from Lung Cancer and Colon Cancer cells induced adipocyte atrophy via NF-κB signaling pathway

doi: 10.1186/s12944-022-01755-2

Figure Lengend Snippet: NF-κB inhibitors partially mitigated lipolysis induced by exosomal IL-8. A The viability of 3 T3-L1 adipocytes treated with different doses of NF-κB inhibitor BAY11-7082 as assessed by CCK8. B-D Western blot and the densitometric analysis of phosphorylated-p65, total p65, UCP1, and PGC1α in adipocytes co-cultured with IL-8 (with and without BAY11-7082), LLC-EVs and/or C26-EVs (with and without BAY11-7082). The blot shown were representatives of three independent experiments. 10 μg EVs were used in this experiment. E Oil red O staining was assessed in the above groups. Bar = 100 μm. F Relative levels of glycerol release in cell supernatants with the above treatments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Reagents and sources: recombinant mouse CXCL15/Lungkine protein (442-LK-002, R&D Systems); anti-IL-8 (AF-208-SP, R&D Systems); CXCR2 inhibitor (SB225002, MCE); NF-κB inhibitor (BAY 11-7082, Sigma-Aldrich); Oil red O solution (Servicebio, China).

Techniques: Western Blot, Cell Culture, Staining