Journal: Cell

Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling

doi: 10.1016/j.cell.2017.02.011

Figure Lengend Snippet: (A) Ex vivo CFU-GM colony formation assay. Sorted mouse LSK HSPCs were cultured in semi-solid methylcellulose medium with 100 ng/ml recombinant mouse IL-3 and the indicated concentration of SCF variants. The number of CFU-GM colonies were scored after a 7-day period and normalized to the top data point in the 5000 ng/ml SCF group. (B) Ex vivo proliferation of purified LSK HSPCs from mouse bone marrow. Cells were cultured on irradiated OP9 stromal cells for 6 days in the presence of titrating concentrations of SCF variants, and the total number of hematopoietic cells generated was quantified. (C) β-hexosaminidase release by human PBCMCs treated with anti-IgE alone or with anti-IgE together with SCF or S4-3a at indicated concentrations (ng/ml) for 30 min in vitro. See also Figure S4. (D) Dose-response curves of IL-6 release by mouse BMCMCs stimulated for 6 h with SCF variants in vitro. (A and C) Data represent mean ± SEM and are pooled from at least two independent experiments with a total of n 6. *p<0.05, **p<0.01, ***p<0.001, and ns = not significant (i.e., p>0.05) compared to the PBS-treated control group by unpaired, two-tailed Student’s t test. (B and D) Data represent mean ± SEM and are representative of at least two independent experiments with n = 3 per experiment. The vertical dotted lines indicate the EC50 values of the dose-response curves. The numbers on top of the black arrows indicate the fold difference between the EC50s.

Article Snippet: Proliferation of HSPCs ex vivo 5000 mouse LSK HSPCs from GFP + C57B/6J mice were plated on irradiated (2000 Rads) OP9 stroma (ATCC) in Alpha MEM media, supplemented with 20% FBS, 1% Pen Strep Glut (Gibco), and either SCF or S4-3a at 5000, 500, 50 and 5 ng/ml.

Techniques: Ex Vivo, Colony Assay, Cell Culture, Recombinant, Concentration Assay, Purification, Irradiation, Generated, In Vitro, Control, Two Tailed Test

Journal: Cell

Article Title: Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling

doi: 10.1016/j.cell.2017.02.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Proliferation of HSPCs ex vivo 5000 mouse LSK HSPCs from GFP + C57B/6J mice were plated on irradiated (2000 Rads) OP9 stroma (ATCC) in Alpha MEM media, supplemented with 20% FBS, 1% Pen Strep Glut (Gibco), and either SCF or S4-3a at 5000, 500, 50 and 5 ng/ml.

Techniques: Recombinant, Mutagenesis, Electron Microscopy, Staining, Modification, Giemsa Stain, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Endotoxin Assay, Selection, Bacteria, Expressing, Sequencing, Software, Diagnostic Assay, Live Cell Imaging