Journal: Respiratory Research

Article Title: Melatonin improves influenza virus infection-induced acute exacerbation of COPD by suppressing macrophage M1 polarization and apoptosis

doi: 10.1186/s12931-024-02815-0

Figure Lengend Snippet: The effects of melatonin on abnormal polarization of pulmonary macrophages in AECOPD. a , b Mouse lung tissue sections from each group were probed with the specific antibody against pulmonary macrophage marker F4/80 (red) and co-probed with antibodies against M1 marker (iNOS) or M2 marker (CD206), and, representative lung immunofluorescence staining of lung tissue sections were shown (original magnification × 100 and × 500). c , d Western blot analysis of the expression of total-STAT1, phospho-STAT1, total-STAT6, phospho-STAT6, iNOS and Arg1 to β-tublin in lung tissue homogenates from air group, COPD (CS exposure) group, AECOPD (smoke + H3N2) group, AECOPD + melatonin (Mel, 30 mg/kg) group. Data expressed as mean ± SD ( n ≥ 3). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The membranes were incubated with primary antibodies: total-STAT1 (1:1000, PTM-5754, PTM Bio, Hangzhou, China), Phospho-STAT1 (1:1000, 340797, ZenBio, Chengdu, China), total-STAT6 (1:1000, 380957, ZenBio), Phospho-STAT6 (1:500, sc-136019, Santa Cruz), iNOS (1:1000, ab178945, Abcam), Arg1 (1:1000, #93668S, Cell signaling technology), Caspase1 (1:500, sc-392736, Santa Cruz), MT1/2 (1:500, sc-398788, Santa Cruz), β-Tublin (1:5000, M20005, Abmart, Shanghai, China) and GAPDH (1:5000, ab181602, Abcam) overnight at 4℃.

Techniques: Marker, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Respiratory Research

Article Title: Melatonin improves influenza virus infection-induced acute exacerbation of COPD by suppressing macrophage M1 polarization and apoptosis

doi: 10.1186/s12931-024-02815-0

Figure Lengend Snippet: The effects of melatonin abnormal polarization of IAV-infected CSE-stimulated macrophages. a - c Western blot analysis of the expression of total-STAT1, Phospho-STAT1, total-STAT6, Phospho-STAT6 and IAV-NP to GAPDH as well as iNOS, Arg1 and MT1/2 to GAPDH in CSE-stimulated Raw264.7 cells infected by IAV/H3N2 infection (MOI = 2, 12 h) with/without melatonin pretreatment (10 μM, 100 μM and 200 μM, 3 h before H3N2 infection). d Representative Immunofluorescence images of iNOS (green) and Arg1 (red) expression in Raw264.7 cells infected by H3N2 infection (MOI = 2, 12 h) with/without melatonin pretreatment (200 μM, 3 h before H3N2 infection) (bar = 10 μm, original magnification × 630). e Quantitative RT-PCR measurements of the relative mRNA levels of MCP1, TNF-α, Arg1 and Fizz1 in CSE-stimulated Raw264.7 cells. Data expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.001

Article Snippet: The membranes were incubated with primary antibodies: total-STAT1 (1:1000, PTM-5754, PTM Bio, Hangzhou, China), Phospho-STAT1 (1:1000, 340797, ZenBio, Chengdu, China), total-STAT6 (1:1000, 380957, ZenBio), Phospho-STAT6 (1:500, sc-136019, Santa Cruz), iNOS (1:1000, ab178945, Abcam), Arg1 (1:1000, #93668S, Cell signaling technology), Caspase1 (1:500, sc-392736, Santa Cruz), MT1/2 (1:500, sc-398788, Santa Cruz), β-Tublin (1:5000, M20005, Abmart, Shanghai, China) and GAPDH (1:5000, ab181602, Abcam) overnight at 4℃.

Techniques: Infection, Western Blot, Expressing, Immunofluorescence, Quantitative RT-PCR