Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition reduced lung tissues damage induced by lung I/R (A) The Gal3 protein expression in lung tissue was assessed by western blotting, and the density of Gal3 (relative to β-actin) on western blotting was quantified (n = 3). (B–D) The expression of Gal3 in lung tissue, BALF and serum was detected by ELISA (n = 6). (E) Paraffin-embedded lung tissue sections were stained with hematoxylin and eosin (n = 3) at magnifications of 200×, scale bar 200 μm and 400X, scale bar 100 μm. (F) The extent of lung injury was assessed by scoring the F images (n = 6 section per group). (G) The W/D ratio of lung tissue (n = 6). (H) Ultrastructural changes were evaluated through transmission electron microscopy. Arrows indicate lamellar body and triangles mitochondria (n = 3). Scale bar, 1 μm. The data were mean ± SEM. One-way analysis of variance was used for data comparison, ns, not significant, ∗p < 0.05, was statistically significant.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Transmission Assay, Electron Microscopy, Comparison

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition attenuated the inflammatory response induced by lung I/R (A–C) BALF of left lung after lung I/R was collected, and the supernatant was extracted to detect the contents of TNF-α, IL-1β, and IL-10 in BALF. (D–F) After LIRI, the heart blood was taken, and the supernatant was collected by centrifugation after standing at low temperature for 3 h. The levels of TNF-α, IL-1β, and IL-10 in the serum were detected. (G–I) The lung tissue was subjected to standard treatment to detect the contents of TNF-α, IL-1β, and IL-10 in lung tissue. Data shown are mean ± SEM from n = 6 per group, ns, no statistical significance, ∗p < 0.05, was statistically significant.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Centrifugation

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Inhibition of Gal3 reduces the expression of M1-type macrophages induced by lung I/R (A) Western blot of IL-1β, IL-6, TNF-α, and CCL5 in lung tissues. (B–E) Density analysis of IL-1β, IL-6, TNF-α, and CCL5 relative to β-actin in part A. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Lung tissues were stained with macrophages marker (F4/80, green), anti-IL-1β (red), and anti-IL-6 antibodies (red). Nuclei were stained with DAPI (blue). Magnification, 400×, scale bar, 100 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

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

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Inhibition of Gal3 increases the expression of M2 type macrophages induced by lung I/R (A) Western blot of Arg1, IL-10, Retnla, and Chil3 in lung tissues. (B–E) Density analysis of Arg1, IL-10, Retnla, and Chil3 relative to β-actin in part A. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Lung tissues were stained with macrophages marker (F4/80, green), anti-Arg1 (red), and anti-IL-10 antibodies (red). Nuclei were stained with DAPI (blue). Magnification, 400×, scale bar, 100 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

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

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition reduces macrophage polarization toward M1-type in OGD/R vitro model (A) Western blot of Gal3 in lung tissues. (B) Density analysis of Gal3 relative to β-actin in part A. (C) Cell viability was measured by CCK-8 assay (n = 6). (D) Western blot of IL-1β, IL-6, TNF-α, and CCL5 in RAW264.7 cells. (E–H) Densitometry of western blots in part D. Levels were standardized uniformly with β-actin. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (I and J) Immunofluorescence staining showed macrophages marker (F4/80, green), IL-1β (red), and IL-6 (red) in RAW264.7 cells. Nuclei were stained with DAPI (blue). Magnification, 200×, scale bar, 200 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Western Blot, CCK-8 Assay, Immunofluorescence, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition promotes the polarization of macrophages toward the M2-type in OGD/R vitro model (A) Western blot of Arg1, IL-10, Retnla, and Chil3 in RAW264.7 cells. (B–E) Densitometry of western blots in part A. Levels were standardized uniformly with β-actin. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Immunofluorescence staining showed macrophages marker (F4/80, green), Arg1 (red), and IL-10 (red) in RAW264.7 cells. Nuclei were stained with DAPI (blue). Magnification, 200×, scale bar, 200 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

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

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 and TREM2 colocalized in macrophages and Gal3 inhibited the expression of TREM2 in LIRI (A) Macrophage markers (F4/80, green), anti-TREM2 antibody (red), and anti-Gal3 antibody (pink) were used for immunofluorescence co-localization staining of lung tissue. Magnification, 400×, scale bar, 200 μm. (B) Enlarge the dotted box in part A. F4/80 (green), TREM2 (red), and Gal3 (pink) co-localization qualitative analysis. White dots to white triangles area (white dotted line) were analyzed with line intensity scans at higher magnification. Measurement of the intensity values demonstrated co-localization in those regions. Magnification, 2000×, scale bar, 50 μm. (C) Western blotting and (E) RT-qPCR expression of TREM2 in lung tissue. (D) Western blotting and (F) RT-qPCR expression of TREM2 in RAW264.7 cells. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05).

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

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

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet:

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Recombinant, Modification, Saline, Protease Inhibitor, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Targeting fibroblast CD248 attenuates CCL17-expressing macrophages and tissue fibrosis

doi: 10.1038/s41598-020-73194-x

Figure Lengend Snippet: CD248 interacted with galectin-3 to induce CCL17-expressing pro-fibrotic macrophages. ( a ) qPCR of genes encoding cytokines and chemokines in macrophages isolated from D7 UUO kidneys of WT and Lgals3 knockout ( Lgals3 –/– ) mice. n = 5. ( b,c ) HEK293T cells were transfected with and without plasmid DNA expressing galectin-3 or CD248-DDK separately. CD248-DDK (DDK-IP) or galectin-3 (Gal3-IP) was immunoprecipitated from the cell lysates, and then immunoblot analyses of galectin-3 and CD248-DDK were performed. ( d ) Flow cytometry of the binding of CD248 extracellular domain (CD248ECD-EGFP) to WT and Lgals3 –/– BMDMs. Blue and red lines indicate results for WT and Lgals3 –/– BMDMs, respectively, with CD248ECD-EGFP. Black and gray lines indicate results for WT and Lgals3 –/– BMDMs, respectively, with control medium. ( e ) Flow cytometry of CD248ECD-EGFP binding to WT BMDMs in the presence of 25 mM lactose (red) or sucrose (blue). Conditioned medium from HEK293T cells in the presence of lactose (gray) or sucrose (black) was used as control. ( f ) qPCR of Ccl17 in WT and Lgals3 –/– BMDMs in the absence (control) or presence of 200 ng/ml rCD248 for 48 h. n = 6. ( g ) Gel plots of PCR for Ccr4 and Gapdh in D7 UUO-kidney myofibroblasts isolated from WT and Cd248 –/– mice. The reaction cycles for Ccr4 and Gapdh were 30 and 15, respectively. ( h ) qPCR for genes Col1a1 and Acta2 in isolated D7 UUO-kidney myofibroblasts after incubation with vehicle (Con), TGF-β1, and CCL17 for 24 h. Data are expressed as means ± standard errors of the mean. * P < 0.05, ** P < 0.01 and *** P < 0.001 by unpaired t-test in ( a ) and one-way ANOVA with post hoc Tukey’s multiple comparisons test in ( f , h ).

Article Snippet: HEK293T cells were transfected with plasmid pCMV6- Cd248 -DDK (MR210537; OriGene Technologies, Inc., Rockville, MD) or pCMV6- Lgals3 (MC208879; OriGene Technologies, Inc.) by PolyJet reagent (SignaGen Laboratories, Rockville, MD).

Techniques: Expressing, Isolation, Knock-Out, Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Flow Cytometry, Binding Assay, Incubation