Journal: STAR Protocols

Article Title: Generation of hematopoietic lineage cell-specific chimeric mice using retrovirus-transduced fetal liver cells

doi: 10.1016/j.xpro.2022.101526

Figure Lengend Snippet:

Article Snippet: Mouse SCF, research grade , Miltenyi Biotec , 130-101-741.

Techniques: Virus, Recombinant, Marker, Modification, Saline, Transfection, Western Blot, Plasmid Preparation, Purification, Gel Extraction, Retroviral, Sequencing, Software, Flow Cytometry

Journal: Autophagy

Article Title: Kit-mediated autophagy suppression driven by a viral oncoprotein emerges as a crucial survival mechanism in Merkel cell carcinoma

doi: 10.1080/15548627.2025.2477385

Figure Lengend Snippet: MCPyV truncated LT induces paranuclear retention and stabilization of KIT. (A) Immunofluorescence detection of KIT (green) and MCPyV LT (CM2B4, red) or sT (CM8E6, red) in KIT-HEK293 cells transfected with MCPyV T-antigens or vector control ( n = 6). (B) Top : illustration of the expression constructs of LT339 and the VPS39-interaction defective mutant LT339 W209A . Bottom : Representative images showing the effect of LT339 and LT339 W209A mutant on localization of KIT (green). LT was detected by CM2B4 (red). ( n = 3) (C) Immunoblots showing the effect of the LT339 and LT339 W209A on KIT expression. The quantification of KIT level is shown below the immunoblots ( n = 9). (D) Top : immunoblot analysis of the effect LT339 and LT339 W209A on KIT protein stability in the presence of cycloheximide (CHX) up to 4 h. Bottom : quantification of KIT protein stability after normalization to 0 h time point ( n = 4). (E) Immunoblots showing KIT protein stability in KIT-HEK293 cells transfected with LT339, LT339 W209A or plasmid control (CTR) treated with KIT ligand (KITLG, 100 ng/mL) or solvent control (PBS) in the presence of CHX (100 μg/mL). (F) Quantification of KIT protein stability after normalization to 0 h time point ( n = 5). Solid lines represent PBS control (KITLG − ) and dotted lines represent KITLG treatment (KITLG + ). (A and B) Nuclei were stained by DAPI (blue). Scale bar: 10 μm. Numbers below the images refer to the proportion of cells with KIT paranuclear dot-like staining to the total number of cells analyzed. (C, D and F) Error bars represent mean ± SEM. * p < 0.05, *** p < 0.001, ns = not significant were calculated by one-way ANOVA with post-hoc Tukey’s test (C), two-way ANOVA (D) or two-way ANOVA with post-hoc Bonferroni’s test (F).

Article Snippet: For KITLG induced KIT degradation experiments, KIT-293 cells were transfected with LT339, LT339 W209A or CTR for 48 h, followed by addition of CHX (100 μg/mL) and KITLG (100 ng/mL; Proteintech Group, HZ-1024) or PBS.

Techniques: Immunofluorescence, Transfection, Plasmid Preparation, Control, Expressing, Construct, Mutagenesis, Western Blot, Solvent, Staining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: The stem cell factor-c-kit system and mast cells in human pancreatic cancer.

doi: 10.1097/01.lab.0000036875.21209.f9

Figure Lengend Snippet: Figure 1. A and B, Stem cell factor (SCF) and c-kit immunostaining in normal pancreatic tissues. A duct showing hyperplastic features exhibits strong c-kit-immunoreactivity (A) and it is surrounded by c-kit–positive (A) and SCF-positive (B) inflammatory cells. C, Immunostaining performed with the highly specific anti-tryptase antibody reveals that most of the inflammatory cells described around large and hyperplastic ducts are mast cells (C). Scale bar 25 m.

Article Snippet: The following materials were purchased: FCS, RPMI and DMEM, trypsin-EDTA solution, and penicillinstreptomycin solution from Biochrom KG (Berlin, Germany); human recombinant SCF from R&D Systems (Abingdon, United Kingdom) and from Amgen GmbH (Munich, Germany); anti-SCF antibody from Santa Cruz Biotechnology (Santa Cruz, California); anti-c-kit polyclonal antibodies from Santa Cruz Biotechnology and from NeoMarkers (Fremont, California); antitryptase and anti-chymase monoclonal antibodies from Chemicon International (Temecula, California); biotinylated secondary antibodies and streptavidin/ peroxidase complex from Kierkegaard and Perry Laboratories (Gaithersburg, Maryland); FITC-conjugated anti-rabbit antibody from Zymed (San Francisco, California); OCT compound from Miles Inc. (Elkhart, Indiana); horseradish peroxidase-conjugated secondary antibodies and ECL Western blotting detection reagents from Amersham Life Science (Amersham, United Kingdom); and Protease Inhibitor Cocktail Tablets from Roche (Basel, Switzerland).

Techniques: Immunostaining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: The stem cell factor-c-kit system and mast cells in human pancreatic cancer.

doi: 10.1097/01.lab.0000036875.21209.f9

Figure Lengend Snippet: Figure 4. Western blot analysis of SCF, c-kit, and tryptase in normal pancreas and pancreatic cancer samples, as described in “Materials and Methods.” The highly specific anti-SCF antibody detected a single band of approximately 45 kDa in the cancer samples. The anti-c-kit antibody detected one band of approximately 80 kDa in two of the cancer samples, and a band of about 125 kDa in a third cancer sample. The anti-tryptase antibody detected two bands of approximately 30 and 35 kDa in two pancreatic cancer samples, and a single, intermediate-sized band in one cancer and one normal pancreas sample.

Article Snippet: The following materials were purchased: FCS, RPMI and DMEM, trypsin-EDTA solution, and penicillinstreptomycin solution from Biochrom KG (Berlin, Germany); human recombinant SCF from R&D Systems (Abingdon, United Kingdom) and from Amgen GmbH (Munich, Germany); anti-SCF antibody from Santa Cruz Biotechnology (Santa Cruz, California); anti-c-kit polyclonal antibodies from Santa Cruz Biotechnology and from NeoMarkers (Fremont, California); antitryptase and anti-chymase monoclonal antibodies from Chemicon International (Temecula, California); biotinylated secondary antibodies and streptavidin/ peroxidase complex from Kierkegaard and Perry Laboratories (Gaithersburg, Maryland); FITC-conjugated anti-rabbit antibody from Zymed (San Francisco, California); OCT compound from Miles Inc. (Elkhart, Indiana); horseradish peroxidase-conjugated secondary antibodies and ECL Western blotting detection reagents from Amersham Life Science (Amersham, United Kingdom); and Protease Inhibitor Cocktail Tablets from Roche (Basel, Switzerland).

Techniques: Western Blot

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: The stem cell factor-c-kit system and mast cells in human pancreatic cancer.

doi: 10.1097/01.lab.0000036875.21209.f9

Figure Lengend Snippet: Figure 5. Western blot analysis of SCF and c-kit in TAKA-1 cells and in six pancreatic cancer cell lines. TAKA cells did not express SCF, whereas SCF was expressed by Bx-PC-3, MiaPaCa-2, and T3M4 cell lines, the most intense band being present in MiaPaCa-2 cells. The anti-c-kit antibody detected two bands of approximately 80 kDa and one band of 125 kDa in TAKA cells and two bands of approximately 80 kDa in all the cancer cell lines; Capan-1 showed the highest c-kit expression levels.

Article Snippet: The following materials were purchased: FCS, RPMI and DMEM, trypsin-EDTA solution, and penicillinstreptomycin solution from Biochrom KG (Berlin, Germany); human recombinant SCF from R&D Systems (Abingdon, United Kingdom) and from Amgen GmbH (Munich, Germany); anti-SCF antibody from Santa Cruz Biotechnology (Santa Cruz, California); anti-c-kit polyclonal antibodies from Santa Cruz Biotechnology and from NeoMarkers (Fremont, California); antitryptase and anti-chymase monoclonal antibodies from Chemicon International (Temecula, California); biotinylated secondary antibodies and streptavidin/ peroxidase complex from Kierkegaard and Perry Laboratories (Gaithersburg, Maryland); FITC-conjugated anti-rabbit antibody from Zymed (San Francisco, California); OCT compound from Miles Inc. (Elkhart, Indiana); horseradish peroxidase-conjugated secondary antibodies and ECL Western blotting detection reagents from Amersham Life Science (Amersham, United Kingdom); and Protease Inhibitor Cocktail Tablets from Roche (Basel, Switzerland).

Techniques: Western Blot, Expressing

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: The stem cell factor-c-kit system and mast cells in human pancreatic cancer.

doi: 10.1097/01.lab.0000036875.21209.f9

Figure Lengend Snippet: Figure 8. Specificity of the effect of SCF on TAKA-1 cells. TAKA-1 cells were incubated in the absence or presence of SCF (100 ng/ml), anti-c-kit antibody (5 g/ml), or the combination of both reagents. MTT assay was performed after 72 hours. Percent growth inhibition was determined by comparison with control cell growth. * p 0.05.

Article Snippet: The following materials were purchased: FCS, RPMI and DMEM, trypsin-EDTA solution, and penicillinstreptomycin solution from Biochrom KG (Berlin, Germany); human recombinant SCF from R&D Systems (Abingdon, United Kingdom) and from Amgen GmbH (Munich, Germany); anti-SCF antibody from Santa Cruz Biotechnology (Santa Cruz, California); anti-c-kit polyclonal antibodies from Santa Cruz Biotechnology and from NeoMarkers (Fremont, California); antitryptase and anti-chymase monoclonal antibodies from Chemicon International (Temecula, California); biotinylated secondary antibodies and streptavidin/ peroxidase complex from Kierkegaard and Perry Laboratories (Gaithersburg, Maryland); FITC-conjugated anti-rabbit antibody from Zymed (San Francisco, California); OCT compound from Miles Inc. (Elkhart, Indiana); horseradish peroxidase-conjugated secondary antibodies and ECL Western blotting detection reagents from Amersham Life Science (Amersham, United Kingdom); and Protease Inhibitor Cocktail Tablets from Roche (Basel, Switzerland).

Techniques: Incubation, MTT Assay, Inhibition, Comparison, Control