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

Journal: PLoS ONE

Article Title: Cytomegalovirus pp71 Protein Is Expressed in Human Glioblastoma and Promotes Pro-Angiogenic Signaling by Activation of Stem Cell Factor

doi: 10.1371/journal.pone.0068176

Figure Lengend Snippet: A : HUVECs stimulated with either recombinant human SCF or conditioned medium from NPCs transduced with rAD-GFP, rAD-pp71, or rAD-pp71+ SCF neutralizing antibody were analyzed by western blot for phosphor-c-Kit, total c-kit, phosphor-p38, total p38, and actin. B : HUVECs were grown overnight in gel matrix supplemented as indicated. Capillary tubes that formed in each condition were visualized by microscopy (top panel), counted and plotted (bottom panel). *p = <0.05 as determined by student t-test when rAD-pp71 was compared to rAD-GFP and when rAD-pp71+Nab was compared to rAD-pp71. Each condition was run in duplicate and the experiment was repeated three times. C : HUVEC cells were subjected to a 4 h transwell migration assay using either 20,000 U87 cells treated as indicated (dark grey bars) or supernatant obtained from the same cells (light grey bars) on the bottom wells. For negative and positive control we used serum-free medium or rhSCF (1 ug/mL) respectively. HUVECs that migrated to the bottom of the well were stained and counted. *p = 0.015 for cells and 0.032 for supernatant when compared to untreated control, or 0.058 and 0.075 when compared to rAD-GFP, respectively by student t-test. Each condition was run in triplicate and the experiment was repeated twice. D : Frozen GBM tissue sections were immunostained for pp71 alone or co-immunostained for pp71 (green) and the endothelial cell marker CD31 (red). Tissues were counterstained with DAPI. Two regions from a representative example from a GBM sample are shown. Magnification of the region of interest is shown in the last panel. Bar = 200µm. Negative controls can be found in .

Article Snippet: Fresh supernatants from NPCs or U87 cells were harvested, and 20 ng/mL recombinant human SCF (R&D Systems) was incubated with untreated supernatant and 1 ug/mL of anti-SCF neutralizing antibody (R&D Systems) was incubated with rAD-pp71 supernatant for 1 hour at 37 degrees.

Techniques: Recombinant, Transduction, Western Blot, Microscopy, Transwell Migration Assay, Positive Control, Staining, Marker