Journal: iScience

Article Title: Off-the-shelf third-party HSC-engineered iNKT cells for ameliorating GvHD while preserving GvL effect in the treatment of blood cancers

doi: 10.1016/j.isci.2022.104859

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Article Snippet: Recombinant human IL-3 , Peprotech , CAT#200–03.

Techniques: Enzyme-linked Immunosorbent Assay, Purification, Control, Blocking Assay, Virus, Recombinant, Cell Culture, Saline, Plasmid Preparation, Software, Imaging

Journal:

Article Title: Involvement of Prolonged Ras Activation in Thrombopoietin-Induced Megakaryocytic Differentiation of a Human Factor-Dependent Hematopoietic Cell Line

doi:

Figure Lengend Snippet: (A) Flow cytometric analysis of F-36P and F-36P-mpl cells. The expression of c-Mpl was examined by staining with rabbit anti-c-Mpl serum (——) or preimmune rabbit serum (–––). (B) Dose response of F-36P and F-36P-mpl to rhIL-3 and rhTPO. Triplicate aliquots of cells were cultured in serum-free medium with various concentration of rhIL-3 or rhTPO for 48 h, and then cell proliferation was measured by a [3H]thymidine incorporation assay (F-36P; □, IL-3; ○, TPO; F-36P-mpl; ▪, IL-3; •, TPO). (C) Changes in the total viable cell number during culture with or without rhIL-3 or rhTPO. F-36P-mpl cells were resuspended in 10% fetal calf serum–RPMI containing 10-ng/ml rhIL-3 or 30-ng/ml rhTPO at a cell density of 100/μl, and the total viable cell number was determined by the trypan blue dye exclusion method at the time indicated (□, IL-3; ○, TPO; ▵, free of growth factor). The results shown are the means ± the standard deviations of triplicate cultures.

Article Snippet: Highly purified recombinant human TPO (rhTPO) and rhIL-3 were provided by the Kirin Brewery Company Ltd. (Tokyo, Japan).

Techniques: Expressing, Staining, Cell Culture, Concentration Assay, Thymidine Incorporation Assay

Journal:

Article Title: Involvement of Prolonged Ras Activation in Thrombopoietin-Induced Megakaryocytic Differentiation of a Human Factor-Dependent Hematopoietic Cell Line

doi:

Figure Lengend Snippet: TPO-induced tyrosine phosphorylation of STAT proteins in F-36P-mpl cells. F-36P-mpl cells were serum and factor starved for 12 h and then either left unstimulated or stimulated with rhTPO for 15 min. Total cell lysates were immunoprecipitated with an anti-STAT1, an anti-STAT3, or an anti-STAT5b Ab. The blots were probed with an antiphosphotyrosine (α-PY) MAb. The filters were then stripped and reprobed with anti-STAT1, anti-STAT3, and anti-STAT5b Abs, respectively.

Article Snippet: Highly purified recombinant human TPO (rhTPO) and rhIL-3 were provided by the Kirin Brewery Company Ltd. (Tokyo, Japan).

Techniques: Immunoprecipitation

Journal:

Article Title: Involvement of Prolonged Ras Activation in Thrombopoietin-Induced Megakaryocytic Differentiation of a Human Factor-Dependent Hematopoietic Cell Line

doi:

Figure Lengend Snippet: Effects of dn STATs and dn ras on rhTPO- and rhIL-3-induced proliferation. Cells of each clone were IL-3 starved for 24 h and then cultured for 48 h with rhTPO or rhIL-3 at the concentrations indicated. The cells were left untreated or treated with 0.5 mM IPTG during the 24-h starvation period and the 48-h culture period. Cell proliferation was quantitated with a [3H]thymidine incorporation assay. The results shown are means ± the standard deviations of triplicate experiments. □, IL-3 without IPTG; ▪, IL-3 with IPTG; ○, TPO without IPTG; •, TPO with IPTG.

Article Snippet: Highly purified recombinant human TPO (rhTPO) and rhIL-3 were provided by the Kirin Brewery Company Ltd. (Tokyo, Japan).

Techniques: Cell Culture, Thymidine Incorporation Assay