Journal: STAR Protocols

Article Title: Dynamic BH3 profiling method for rapid identification of active therapy in BH3 mimetics resistant xenograft mouse models

doi: 10.1016/j.xpro.2021.100461

Figure Lengend Snippet:

Article Snippet: Phosal 50 PG , Fisher Scientific , Cat# NC0130871.

Techniques: Blocking Assay, Recombinant, Electron Microscopy, Red Blood Cell Lysis, Software, Cell Culture, Binding Assay, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Venetoclax or Ruxolitinib in Pre-Transplant Conditioning Lowers the Engraftment Barrier by Different Mechanisms in Allogeneic Stem Cell Transplant Recipients

doi: 10.3389/fimmu.2021.749094

Figure Lengend Snippet: Donor cell engraftment is dependent on recipient pre-alloSCT irradiation dose and treatment with BCL2 or JAK1/2 inhibitors. WT mice were treated with venetoclax or ruxolitinib, or their respective vehicle for two days. The following day mice were treated with RIC and alloSCT. (A) Donor cell engraftment (H2kd+ cells) was measured in the blood at day 21 post-alloSCT. WT mice were treated with venetoclax or ruxolitinib, or their respective vehicle for two days. Mice (n=3-4/group) were killed on days 1, 2, 3, and 7, and BM was harvested and analysed by flow cytometry for the absolute number of (B–F) NK cells (NK1.1+CD3-), cNK (NKp46+CD49b+), M1 mature (CD11b+CD27+), M2 mature (CD11b+CD27-) and immature (CD11b-CD27+) NK cells; (G–L) naive (N; CD44-CD62L+); central memory (CM; CD44+CD62L+); effector memory (EM; CD44+CD62L-) CD4+ and CD8+ T cells. Data is representative of 3 independent experiments. Statistical analysis was performed using unpaired T test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Venetoclax (100 mg/kg) and its vehicle (60% phosal R 50 PG (Merck, Germany), 30% polyethylene glycol (PEG) 400 (Merck, Germany), 10% ethanol) were administered by oral gavage once daily for two days, with a cumulative total dose of 4 mg. Ruxolitinib (180 mg/kg) and its vehicle (2% DMSO, 30% PEG 300 (Merck, Germany), ddH2O) were administered twice a day by oral gavage for two days, with a cumulative total dose of 14.4 mg.

Techniques: Irradiation, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Venetoclax or Ruxolitinib in Pre-Transplant Conditioning Lowers the Engraftment Barrier by Different Mechanisms in Allogeneic Stem Cell Transplant Recipients

doi: 10.3389/fimmu.2021.749094

Figure Lengend Snippet: Venetoclax and Ruxolitinib differentially affect MHC class-II and IFN-inducible gene expression. WT mice were treated with venetoclax or ruxolitinib, or their respective vehicle for two days. Mice (n=3-4/group) were killed on days 1, 3, and 7, and gene expression was determined from BM RNA using the NanoString Mouse PanCancer Immune Profiling Panel. (A) Heat map of relative gene expression of H2-DMb2, H2-Ab1, H2-Eb1, H2-Aa, H2-Ob, CD74, Tap1, Rsad2, Klrb1 and Isg15 from venetoclax and ruxolitinib-treated mice. Relative expression of Rsad2, Klrb1, H2-DMB2 and CD74 (B–E) was compared between venetoclax or ruxolitinib-treated mice and their respective vehicle on day 1 post-treatment. The geometric mean of MHC-II expression on CD19+ B cells (F) , percentage of CD19+ B cells expressing MHC-II (G) , and the fold-change of MHC-II expression on total CD45+ BM cells from venetoclax or ruxolitinib-treated mice was compared to each vehicle control group (H) . Statistical analysis was performed using Mann-Whitney unpaired T test (B–E) , and unpaired T test (F–H) . *p < 0.05, **p < 0.01.

Article Snippet: Venetoclax (100 mg/kg) and its vehicle (60% phosal R 50 PG (Merck, Germany), 30% polyethylene glycol (PEG) 400 (Merck, Germany), 10% ethanol) were administered by oral gavage once daily for two days, with a cumulative total dose of 4 mg. Ruxolitinib (180 mg/kg) and its vehicle (2% DMSO, 30% PEG 300 (Merck, Germany), ddH2O) were administered twice a day by oral gavage for two days, with a cumulative total dose of 14.4 mg.

Techniques: Gene Expression, Expressing, Control, MANN-WHITNEY