lymphocyte antigen 6 complex locus g6d  (Boster Bio)

Journal: Ecotoxicology and environmental safety

Article Title: C5a-C5aR1 axis mediates lung inflammation and fibrosis induced by single-walled carbon nanotubes via promoting neutrophils recruitment.

doi: 10.1016/j.ecoenv.2024.117627

Figure Lengend Snippet: Fig. 3. Early neutrophils recruitment was associated with lung inflammation and fibrosis induced by SWCNT exposure. (A) Flow cytometry analysis of Ly6G+CD11b+ cells in BALF. (B) Quantitative analysis of the proportion of neutrophils in BALF. (C) The content of IL-18 in BALF by ELISA. (D-E) IHC assay and quantification of Ly6G positive cells in lung tissues. Red arrows indicate Ly6G positive cells. The correlation of the proportion of neutrophils (Day 1) and inflam matory score (Day 1) (F) and hydroxyproline content (Day 28) (G) in SWCNT-exposed lung. Data were pooled from at least three independent experiments and expressed as the means ± SD. *P < 0.05 vs CTRL group. Two-tailed unpaired Student’s t-test.

Article Snippet: Next, sections were sealed using normal goat serum and stained with primary antibodies against C5b-9 (Abcam, US), C5a (Abcam, US), C5aR1 (Santa Cruz, US), collagenI (COLI) (Abcam, US), α-smooth muscle actin (α-SMA) (Abcam, US) and lymphocyte antigen 6 complex locus G6D (Ly6G) (Cell Signaling Technology, US), then stained with corresponding secondary antibodies, followed by streptavidin-perosidase, 3,3’-diaminobenzidine (DAB) reactions, and haematoxylin staining.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Ecotoxicology and environmental safety

Article Title: C5a-C5aR1 axis mediates lung inflammation and fibrosis induced by single-walled carbon nanotubes via promoting neutrophils recruitment.

doi: 10.1016/j.ecoenv.2024.117627

Figure Lengend Snippet: Fig. 5. C5a-C5aR1 axis regulated SWCNT-activated neutrophils recruitment. (A) Flow cytometry analysis of Ly6G+CD11b+ cells in BALF. (B) Quantitative analysis of the proportion of neutrophils in BALF. (C) The content of IL-18 in BALF by ELISA. (D-E) IHC assay and quantification of Ly6G positive cells in lung tissues. Red arrows indicate Ly6G positive cells. Data were pooled from at least three independent experiments and expressed as the means ± SD. *P < 0.05 vs CTRL group; #P < 0.05 vs SWCNT group. One-way ANOVA followed by LSD’s multiple comparisons test.

Article Snippet: Next, sections were sealed using normal goat serum and stained with primary antibodies against C5b-9 (Abcam, US), C5a (Abcam, US), C5aR1 (Santa Cruz, US), collagenI (COLI) (Abcam, US), α-smooth muscle actin (α-SMA) (Abcam, US) and lymphocyte antigen 6 complex locus G6D (Ly6G) (Cell Signaling Technology, US), then stained with corresponding secondary antibodies, followed by streptavidin-perosidase, 3,3’-diaminobenzidine (DAB) reactions, and haematoxylin staining.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Science translational medicine

Article Title: A targetable pathway in neutrophils mitigates both arterial and venous thrombosis

doi: 10.1126/scitranslmed.abj7465

Figure Lengend Snippet: (A) In the murine antiphospholipid antibody (aPL)–injected model, mice were injected intraperitoneally (ip) with either aPL isolated from patients with APS (n = 6 mice) or IgG isolated from healthy controls (n = 4 mice) at 24 hours and 1 hour before a carotid artery laser-induced injury model of thrombosis. Time to occlusive thrombus development was measured. (B) Neutrophil depletion in aPL-treated mice using anti-Ly6G antibody (n = 8 mice) or control IgG (n = 8 mice) was followed by carotid artery injury to measure dependency on neutrophils in arterial thrombus formation. Time to occlusive thrombosis development was measured. All mice received aPLs. (C) Neutrophils were harvested from IgG- or aPL-treated wild-type (WT) mice and transferred into untreated WT mice (n = 4 IgG-treated neutrophil recipients; n = 6 aPL-treated neutrophil recipients) before carotid artery injury. (D) Klf2 expression was measured in neutrophils harvested from IgG-injected (n = 3) or aPL-injected (n = 5) WT mice. (E) KLF2 expression was measured in neutrophils harvested from humans with APS (n = 7) or healthy controls (n = 4). *P < 0.05, **P < 0.01, and ***P < 0.001 by unpaired, two-tailed Student’s t test.

Article Snippet: Neutrophil and monocyte depletion For neutrophil depletion studies, mice were injected with anti-Lymphocyte antigen 6 complex locus G6D (Ly6G) antibody (Bio X Cell, clone 1A8) at an initial concentration of 400 μg (injected retro-orbitally) 1 day before thrombosis assays and 100 μg (intraperitoneally) just before surgery.

Techniques: Injection, Isolation, Control, Expressing, Two Tailed Test

Journal: Science translational medicine

Article Title: A targetable pathway in neutrophils mitigates both arterial and venous thrombosis

doi: 10.1126/scitranslmed.abj7465

Figure Lengend Snippet: (A) The volcano plot from RNA sequencing of peripheral blood neutrophils demonstrates more than 2000 differentially expressed genes between K2KO and LysM cells. (B) Gene Ontology (GO) biological process analysis of differentially expressed genes from K2KO neutrophil RNA sequencing is shown. % Gene set represents the proportion of the entire process that is affected by loss of KLF2. (C) The overlap of enriched pathways between K2KO neutrophils and neutrophils from humans with APS is shown. (D) Results of a transwell migration experiment using N-formyl-Met-Leu-Phe (fMLP) as chemoattractant for LysM and K2KO neutrophils are shown (n = 10 to 12). (E) Peritoneal lavage after fMLP intraperitoneal injection was used to quantify neutrophil migration in LysM (n = 7) and K2KO (n = 5) mice. (F and G) Ly6G-stained neutrophils were quantified along vessel walls early after (F) carotid artery injury (10 min) and (G) IVC ligation (4 hours) are shown for LysM and K2KO mice (n = 4 to 5). (H) Neutrophils were quantified along vessel walls early after carotid artery injury in IgG-injected (n = 5) or aPL-injected (n = 4) WT mice. *P < 0.05, **P < 0.01, and ***P < 0.001 by unpaired, two-tailed Student’s t test.

Article Snippet: Neutrophil and monocyte depletion For neutrophil depletion studies, mice were injected with anti-Lymphocyte antigen 6 complex locus G6D (Ly6G) antibody (Bio X Cell, clone 1A8) at an initial concentration of 400 μg (injected retro-orbitally) 1 day before thrombosis assays and 100 μg (intraperitoneally) just before surgery.

Techniques: RNA Sequencing, Migration, Injection, Staining, Ligation, Two Tailed Test

Journal: Science translational medicine

Article Title: A targetable pathway in neutrophils mitigates both arterial and venous thrombosis

doi: 10.1126/scitranslmed.abj7465

Figure Lengend Snippet: (A) Time to occlusive thrombosis was measured in the carotid artery injury model comparing LysM (n = 7) and K2KO (n = 8) mice. (B) IVC thrombus weight measured after IVC ligation comparing LysM and K2KO mice. n = 8 mice. (C) The timeline shows the experimental model of neutrophil depletion studies in K2KO mice. (D and E) Neutrophils were depleted with anti-Ly6G antibody or control antibody in LysM (n = 6 to 7) and K2KO (n = 6 to 8) mice before (D) arterial or (E) venous thrombosis induction. (F) The timeline depicts adoptive transfer experiments wherein neutrophils were isolated from healthy LysM or K2KO mice and infused into LysM/K2KO mice before arterial or venous thrombosis. (G and H) Infusion of LysM (CRE) or K2KO (KO) neutrophils into respective genotypes (n = 6 to 8 per group) was followed by initiation of (G) arterial and (H) venous thrombosis. Data were analyzed using an unpaired, two-tailed Student’s t test.

Article Snippet: Neutrophil and monocyte depletion For neutrophil depletion studies, mice were injected with anti-Lymphocyte antigen 6 complex locus G6D (Ly6G) antibody (Bio X Cell, clone 1A8) at an initial concentration of 400 μg (injected retro-orbitally) 1 day before thrombosis assays and 100 μg (intraperitoneally) just before surgery.

Techniques: Ligation, Control, Adoptive Transfer Assay, Isolation, Two Tailed Test