Project description:This experiment examines how increasing stress (sorbitol) can override normal stemness (NS) conditions, which LIF maintains, causing lineage imbalance and otherwise affecting cells in the early embryo. It also compares NS to normal differentiation (ND) (without LIF). D3 are from 8 blastocysts of 129/Sv+/+ strain mice germ line competent 1. No sex is given as these may actually be a product of different sexes of the 8 blastocysts. The D3 cells were obtained for ATCC (American Type Culture Collection). 1. Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R. The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol 1985;87:27-45. Rex1 promoter- Red fluorescence protein (RFP) transgenic viable reporter ESC were created and validated previously2. 2. Li Q, Gomez-Lopez N, Drewlo S, et al. Development and Validation of a Rex1-RFP Potency Activity Reporter Assay That Quantifies Stress-Forced Potency Loss in Mouse Embryonic Stem Cells. Stem Cells Dev 2016;25:320-8.
Project description:This experiment examines how increasing stress (sorbitol) can override normal stemness (NS) conditions, which LIF maintains, causing lineage imbalance and otherwise affecting cells in the early embryo. It also compares NS to normal differentiation (ND) (without LIF). D3 are from 8 blastocysts of 129/Sv+/+ strain mice germ line competent 1. No sex is given as these may actually be a product of different sexes of the 8 blastocysts. The D3 cells were obtained for ATCC (American Type Culture Collection). 1. Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R. The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol 1985;87:27-45. Rex1 promoter- Red fluorescence protein (RFP) transgenic viable reporter ESC were created and validated previously2. 2. Li Q, Gomez-Lopez N, Drewlo S, et al. Development and Validation of a Rex1-RFP Potency Activity Reporter Assay That Quantifies Stress-Forced Potency Loss in Mouse Embryonic Stem Cells. Stem Cells Dev 2016;25:320-8.
Project description:Smad2, Smad3, Smad4 and Foxh1 ChIPseq performed in pluripotent mESC and embryonic bodies (EBs). RNAseq were performed in WT mESCs and Ebs of WT, Smad2KO, Smad3KO and Smad2/3DKO.
Project description:To dissected the roles of SMAD2/3 and SMAD4 in mouse embryonic stem cells (mESC), knockouts of SMAD2/3 (SMAD2 and SMAD3 double knockout, S2/3DKO) and SMAD4 (SMAD4 knockout, S4KO) were used to conduct RNAseq in ES and EB conditions.
Project description:Human serum derived macrophages were infected with Mtb expressing a transcriptional reporter of viability and on day 5, the cells were sorted for RFP+GFP+ macrophages or RFP+GFP- macrophages.