anti mouse scf neutralizing igg  (R&D Systems)

Journal: Cell death & disease

Article Title: SCF and IL-33 regulate mouse mast cell phenotypic and functional plasticity supporting a pro-inflammatory microenvironment.

doi: 10.1038/s41419-023-06139-7

Figure Lengend Snippet: Fig. 7 SCF controls connective tissue-like MC accumulation in tumor lesions. A Schematic representation of anti-SCF neutralizing Ab administration in vivo. AOM/DSS-treated mice were i.p. injected three times with anti-SCF or control Ab (100 μg/mouse) starting from the fourth DSS cycle. Treated and control mice were sacrificed at 13 weeks from AOM administration. B Colon paraffin-embedded sections from AOM/DSS mice treated with Ctrl-Ig or anti-SCF neutralizing antibodies as described in (A) were stained with anti-MCP4 Ab followed by Alexa Fluor 488 secondary Abs (green). Nuclei were counterstained with DAPI (blue) and images were acquired with a Zeiss LSM980 confocal microscopy using a 20× objective. The frequencies of MCs positive for mMCP4 protease were analyzed in 20 fields randomly acquired from tumor lesions and shown as mean ± SD cells/field. Paired Student’s t test: *p < 0.05. Number of adenomas and colon lengths are shown in (C). Paired Student’s t test: *p < 0.05. Graphs are representative of two independent experiments with a total of 5 mice/group.

Article Snippet: For blocking experiments, mice were i.p. injected with anti-SCF neutralizing antibody (AB-455-NA R&D Systems) or normal goat control IgG (AB-108-C R&D Systems) (100μg/mouse) three times every 10 days starting one day before the fourth DSS cycle (see Fig. 7A).

Techniques: In Vivo, Injection, Control, Staining, Confocal Microscopy

Journal: Scientific Reports

Article Title: Ultraviolet-irradiated endothelial cells secrete stem cell factor and induce epidermal pigmentation

doi: 10.1038/s41598-018-22608-y

Figure Lengend Snippet: Endothelial cells secrete SCF upon UV irradiation: ( a ) Differential gene expression profiling of HDMECs and UV-HDMECs detected by RNA sequencing analysis. N.D. = none detected. ( b ) Expression levels of the mRNA of SCF, EDN1, and TGFβ1 analyzed by real-time PCR. ( c ) SCF protein expression in the UV-HDMECs detected by immunocytochemical staining. Abbreviation: TGF, transforming growth factor; IL, interleukin; IFN, interferon; TNF, tumor necrosis factor; SCF, stem cell factor; EDN, endothelin; FGF, fibroblast growth factor; POMC, proopiomelanocortin; NOS, nitric oxide synthase; KGF, keratinocyte growth factor. ( d ) Levels of secreted SCF in a cultured medium measured by ELISA. The scale bar indicates 100 μm.

Article Snippet: For inhibition studies, melanocytes were treated with a medium conditioned with UVA-irradiated endothelial cells in conjunction with/without KIT-specific inhibitor (ISCK03) or neutralizing anti-SCF antibody (R&D Systems, Minneapolis, MN).

Techniques: Irradiation, Expressing, RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Ultraviolet-irradiated endothelial cells secrete stem cell factor and induce epidermal pigmentation

doi: 10.1038/s41598-018-22608-y

Figure Lengend Snippet: The endothelial cell-derived SCF contributes to skin pigmentation: ( a ) Melanocytes were treated with a KIT-inhibitor, 5 μg/mL ISCK03, in conjunction with a conditioned medium from UV-HDMECs for 3 days. The mRNA (left panel) and protein (right panel) expression levels of MITF and tyrosinase were evaluated using real-time PCR and western blot analysis, respectively. ( b ) A neutralizing anti-SCF antibody (2 μg/mL) was used to block endogenous SCF actions in the conditioned medium from UV-HDMECs. Cropped blots in ( a , b ) were displayed and the full-length blots were shown in Supplementary Fig. . ( c ) SCF (red) and CD31 (green) double-immunostained endothelial cells were detected in the dermis taken from hyperpigmented skin and normal skin from three melasma patients. All values indicate the mean of independent experiments ± SD. The scale bar indicates 200 μm.

Article Snippet: For inhibition studies, melanocytes were treated with a medium conditioned with UVA-irradiated endothelial cells in conjunction with/without KIT-specific inhibitor (ISCK03) or neutralizing anti-SCF antibody (R&D Systems, Minneapolis, MN).

Techniques: Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Blocking Assay