gdf3 mouse 9009 gd 010 r d systems flow cytometry  (R&D Systems)

Journal: Pharmaceuticals

Article Title: GDF3 Protects Mice against Sepsis-Induced Acute Lung Injury by Suppression of Macrophage Pyroptosis

doi: 10.3390/ph17030268

Figure Lengend Snippet: GDF3 protects mice against CLP-Induced ALI. ( a ) Representative images of lung tissue pathology were obtained by performing H&E staining on each group of mice. ( b ) Histological scoring was conducted to assess the injury of CLP-induced ALI. ( c ) A Kaplan–Meier survival curve was generated for each group of mice to compare mortality rates ( n = 10). Values are means ± SEM. *** p < 0.001, **** p < 0.0001.

Article Snippet: We injected recombinant mouse GDF3 protein (20 μg/kg body weight) (R&D Systems, MN, USA) or BSA vehicle into wild-type (WT) mice’s tail vein 3 h before CLP to determine whether it protects against polymicrobial sepsis in vivo.

Techniques: Staining, Generated

Journal: Pharmaceuticals

Article Title: GDF3 Protects Mice against Sepsis-Induced Acute Lung Injury by Suppression of Macrophage Pyroptosis

doi: 10.3390/ph17030268

Figure Lengend Snippet: GDF3 alleviated the increased alveolar–capillary permeability in lung tissue. ( a ) Cells in mouse BALF samples were examined using Wright’s Giemsa staining. ( b ) The total cell counts in BALF samples were measured in CLP-induced ALI mouse models treated with or without GDF3 ( n = 6). ( c ) The W/D ratios of the lung were quantified in mice treated with GDF3 24 h after CLP ( n = 6). Values are means ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: We injected recombinant mouse GDF3 protein (20 μg/kg body weight) (R&D Systems, MN, USA) or BSA vehicle into wild-type (WT) mice’s tail vein 3 h before CLP to determine whether it protects against polymicrobial sepsis in vivo.

Techniques: Permeability, Staining

Journal: Pharmaceuticals

Article Title: GDF3 Protects Mice against Sepsis-Induced Acute Lung Injury by Suppression of Macrophage Pyroptosis

doi: 10.3390/ph17030268

Figure Lengend Snippet: GDF3 inhibits the levels of cytokines in CLP-induced ALI. ( a , b ) Levels of IL-1β, TNF-α in serum ( n = 6). ( c , d ) Levels of IL-1β, TNF-α in BALF ( n = 6). ( e , f ) The mRNA expression levels of IL-1β and TNF-α were assessed in lung tissues. Part of the above cytokines were measured by multiplex secretome analysis ( n = 6). Three independent experiments were conducted. All values are means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: We injected recombinant mouse GDF3 protein (20 μg/kg body weight) (R&D Systems, MN, USA) or BSA vehicle into wild-type (WT) mice’s tail vein 3 h before CLP to determine whether it protects against polymicrobial sepsis in vivo.

Techniques: Expressing, Multiplex Assay

Journal: Pharmaceuticals

Article Title: GDF3 Protects Mice against Sepsis-Induced Acute Lung Injury by Suppression of Macrophage Pyroptosis

doi: 10.3390/ph17030268

Figure Lengend Snippet: GDF3 reduces pyroptosis in CLP-induced ALI. ( a , b ) Caspase-1 and NLRP3 mRNAs were assessed by RT-qPCR (normalized to β-Actin) ( n = 6). ( c , d ) Western blot analysis of Caspase-1 and NLRP3 in the lungs collected from ALI induced by CLP. ( e ) Immunohistochemistry of Caspase-1 and NLRP3 expression in the lung tissues of each group of mice (magnification ×400). ( f , g ) Semi-quantitative analysis of Caspase-1 and NLRP3 expression of lung tissues based on the immunohistochemistry. ( h , i ) The images presented above represent immunofluorescence evidence of Caspase-1 and NLRP3 expression. Blue DAPI staining is applied to the nuclei, F4/80 is stained in red, and Caspase-1 and NLRP3 are stained in green by specific antibodies. Values are means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: We injected recombinant mouse GDF3 protein (20 μg/kg body weight) (R&D Systems, MN, USA) or BSA vehicle into wild-type (WT) mice’s tail vein 3 h before CLP to determine whether it protects against polymicrobial sepsis in vivo.

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

Journal: Pharmaceuticals

Article Title: GDF3 Protects Mice against Sepsis-Induced Acute Lung Injury by Suppression of Macrophage Pyroptosis

doi: 10.3390/ph17030268

Figure Lengend Snippet: GDF3 inhibited LPS-induced pyroptosis in macrophages. ( a – d ) A RT-qPCR was used to measure Caspase-1 and NLRP3 relative mRNA expression levels (normalized to β-Actin) in different macrophages. ( e – g ) Protein expression levels of Caspase-1 and NLRP3 were assessed using Western blotting. Three independent experiments were conducted to obtain the results. Values are means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: We injected recombinant mouse GDF3 protein (20 μg/kg body weight) (R&D Systems, MN, USA) or BSA vehicle into wild-type (WT) mice’s tail vein 3 h before CLP to determine whether it protects against polymicrobial sepsis in vivo.

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Pharmaceuticals

Article Title: GDF3 Protects Mice against Sepsis-Induced Acute Lung Injury by Suppression of Macrophage Pyroptosis

doi: 10.3390/ph17030268

Figure Lengend Snippet: GDF3 reduced the levels of cytokines in LPS-induced macrophages. ( a – d ) TNF-α and IL-1β mRNAs were assessed by RT-qPCR (normalized to β-Actin) in macrophages (Raw264.7 and MHS cells). Values are means ± SEM. * p < 0.05, **** p < 0.0001.

Article Snippet: We injected recombinant mouse GDF3 protein (20 μg/kg body weight) (R&D Systems, MN, USA) or BSA vehicle into wild-type (WT) mice’s tail vein 3 h before CLP to determine whether it protects against polymicrobial sepsis in vivo.

Techniques: Quantitative RT-PCR

Journal: Molecular Neurodegeneration

Article Title: Network-driven plasma proteomics expose molecular changes in the Alzheimer’s brain

doi: 10.1186/s13024-016-0095-2

Figure Lengend Snippet: GDF3 regulates neurogenesis and is reduced in AD brains. a To test whether GDF3 levels are also reduced in AD brains, Human AD and control cortical grey matter regions were lysed and the detergent soluble protein fraction was probed by western blot. Levels of active GDF3 ( b ) were quantified relative to neuron-specific enolase (NSE). c To identify areas in the brain where GDF3 may have a functional role, we referred to The Allen Brain Atlas, which showed strong RNA expression in the mouse hippocampus (blue = high expression). d Using qPCR, GDF3 mRNA expression was detected in non-differentiated adult mouse NPCs and NPCs cultured in differentiating conditions. e To determine whether GDF3 affects stem cell function, adult mouse NPCs were provided recombinant mouse GDF3 and NPC proliferation was assessed using BrdU. f Recombinant mouse GDF3 was also provided to dissociated adult mouse neurospheres and the number of newly formed neurospheres was subsequently quantified. g To investigate whether GDF3 promotes neurogenesis, human-derived NTERA cells stably transfected with Dcx promoter-controlled eGFP were provided recombinant human GDF3. Shown are representative images of DCX-GFP fluorescence expression from an entire well of NTERA cells treated with GDF3 or control for 30 days. h DCX-GFP fluorescence area was quantified relative to the cellular area detected by brightfield microscopy. Results were compared by a one-way ANOVA with a Dunnett's post-test ( b , e ), an unpaired Student’s t test ( d ), or a two-way ANOVA with a Bonferroni post-test ( f , h ) and are representative of at least 2 independent experiments (b n = 16 –18 per group, d,e,f,h; n = 3 per group). Values are mean ± s.e.m.. * p < 0.05, ** p < 0.01, *** p < 0.001 compared to respective control groups

Article Snippet: 5000 cells were plated per well in a 96 well poly-D-lysine/laminin-coated plate with 0, 0.01, 0.1, 1, 10 or 100 ng/ml recombinant mouse GDF3 (R&D Systems) in normal growth media.

Techniques: Control, Western Blot, Functional Assay, RNA Expression, Expressing, Cell Culture, Cell Function Assay, Recombinant, Derivative Assay, Stable Transfection, Transfection, Fluorescence, Microscopy