Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 inhibited LPS-induced apoptosis. AM were isolated from rats and treated with either vehicle (PBS), LPS (to induce ALI), BML-111, BML-111 + LPS. a At 24 h after the treatment, the cell viability was examined by MTT assay. b The apoptosis of cells was determined by flow cytometry following staining the cells with Annexin V and PI. c The expression of different apoptosis biomarkers, including cleaved-Caspase 3, cleaved-Caspase 8,cleaved-Caspase 9, cleaved-PARP, Bcl-2, and Bax was detected by Western blot. Representative Western blot image was shown on the left and the quantification of each protein level relative to that of the internal control (GAPDH) shown on the right. n = 3, * P < 0.05, ** P < 0.01

Article Snippet: AMs were treated with BML-111 for 6 h prior to LPS treatment for a further 2 h. MHY1485 was purchased from MCE (10 μM; MedChem Express, NJ, USA).

Techniques: Isolation, MTT Assay, Flow Cytometry, Staining, Expressing, Western Blot, Control

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 elevated LC3-II level in AMs. AMs were treated with increasing concentrations of BML-111 for 2 h ( a ) or with 100 nM of BML-111 for indicated time periods ( b ). The expression of LC3-I and LC3-II was examined by Western blot. Representative Western blot image was shown on the top and the LC3-II/LC3-I ratio shown on the bottom. n = 3, * P < 0.05, ** P < 0.01

Article Snippet: AMs were treated with BML-111 for 6 h prior to LPS treatment for a further 2 h. MHY1485 was purchased from MCE (10 μM; MedChem Express, NJ, USA).

Techniques: Expressing, Western Blot

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 elevated autophagy level in LPS-treated AM. AMs were treated as indicated. a The expression of LC3-I and LC3-II was examined by Western blot. Representative Western blot image was shown on the top and the LC3-II/LC3-I ratio shown on the bottom. b The expression of LC3-II in AM was detected by immunofluorescence (green signal). All cells were counter stained with DAPI (blue signal). Representative immunofluorescence images from indicated cells were shown on the left and the percentage of LC3-II+ cells quantified and shown as histogram on the right. c The expression of different autophagy and apoptosis biomarkers, including BECN1, SQSTM1/p62 was detected by Western blot. n = 3, * P < 0.05, ** P < 0.01

Article Snippet: AMs were treated with BML-111 for 6 h prior to LPS treatment for a further 2 h. MHY1485 was purchased from MCE (10 μM; MedChem Express, NJ, USA).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 targeted MAPK1 pathway but mTOR-independent mechanism to induce autophagy. a The activation of MAPK1 and MAPK8 was detected by Western blot in AM treated as indicated. Representative Western blot image was shown on the left and the quantification of each protein level relative to that of the internal control (GAPDH) shown on the right. b AM were treated as indicated, in autophagy inhibitor MHY-1485 and mTOR inhibitor Rapamycin. LC3-II expression was examined by immunofluorescence (green signals). All cells were counter stained with DAPI (blue signal). Representative immunofluorescence images from indicated cells were shown on the bottom and the percentage of LC3-II+ cells quantified and shown as histogram on the top. n = 3, * P < 0.05, ** P < 0.01

Article Snippet: AMs were treated with BML-111 for 6 h prior to LPS treatment for a further 2 h. MHY1485 was purchased from MCE (10 μM; MedChem Express, NJ, USA).

Techniques: Activation Assay, Western Blot, Control, Expressing, Immunofluorescence, Staining

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: BML-111 alleviated ALI in vivo. ALI model was established in rats by intratracheal instillation of LPS and rats were either not treated (ALI), or treated with vehicle (PBS + ALI) or BML-111 (BML-111 + ALI). As controls, rats not through ALI induction and treated with either vehicle (PBS) or BML-111 were used. a Upon sacrifice, the lung tissue from each group was examined by HE staining and assessed for ALI score. b The lung tissue was measured for wet/dry weight ratio and compared among different groups. n = 6, * P < 0.05, ** P < 0.01

Article Snippet: AMs were treated with BML-111 for 6 h prior to LPS treatment for a further 2 h. MHY1485 was purchased from MCE (10 μM; MedChem Express, NJ, USA).

Techniques: In Vivo, Staining

Journal: Respiratory Research

Article Title: Lipoxin A4 receptor agonist BML-111 induces autophagy in alveolar macrophages and protects from acute lung injury by activating MAPK signaling

doi: 10.1186/s12931-018-0937-2

Figure Lengend Snippet: The benefits of BML-111 were associated with reduced inflammation and enhanced autophagy in vivo. Bronchoalveolar lavage was collected from rats of each group and the levels of TNF-α ( a ) and IL-6 ( b ) was measured using ELISA. AMs were isolated from rats of each group. c The expressions of TNF-α and IL-6 on the steady-state mRNA level were measured by RT-qPCR. d The expressions of BECN1, SQSTM1/p62, LC3-I, and LC3-II in isolated AM were examined by Western blot. Representative Western blot image was shown on the left and the quantification of each protein level relative to that of the internal control (GAPDH) shown on the right ( e ). n = 6, * P < 0.05, ** P < 0.01

Article Snippet: AMs were treated with BML-111 for 6 h prior to LPS treatment for a further 2 h. MHY1485 was purchased from MCE (10 μM; MedChem Express, NJ, USA).

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Isolation, Quantitative RT-PCR, Western Blot, Control