Prospectively purified bone marrow hematopoietic stem cells (LSK, defined as Lineage- Sca-1+ c-Kit+) and myeloid progenitors (defined as Lineage- Sca-1- c-Kit+) from Mx-Cre-; FoxO1/O3/O4LoxP/LoxP and Mx-Cre+; FoxO1/O3/O4LoxP/LoxP mice 4 weeks after induction of excision with polyinosine-polycyticylic acid (pIpC, 300ug)
Extracted molecule
total RNA
Extraction protocol
Bone marrow hematopoietic stem cells or myeloid progenitors from pooled Mx-Cre-; FoxO1/O3/O4LoxP/LoxP or Mx-Cre+; FoxO1/O3/O4LoxP/LoxP mice were sorted and RNA was prepared using the Qiagen RNeasy micro kit. Amplification and biotin labeling of fragmented amplified cDNA was carried out using the NuGen Ovation Biotin labeling system (NuGEN Technologies, Inc) in triplicate. Labeled probes were hybridized to Affymetrix Mouse Genome 430 2.0 (MOE430_2) GeneChip arrays.
Label
Biotin
Label protocol
NuGen Ovation Biotin labeling system
Hybridization protocol
Affymetrix
Scan protocol
Affymetrix
Description
Study the effect of the deficiency of FoxO1, FoxO3, and FoxO4 in murine bone marrow hematopoietic stem cells and myeloid progenitors on expression of genes involved in reactive oxygen species (ROS) metabolism.
Data processing
Tothova Z, Kollipara R, Huntly BJ, Lee BH, Castrillon DH, Cullen DE, McDowell EP, Lazo-Kallanian S, Williams IR, Sears C, Armstrong SA, Passegue E, DePinho RA, Gilliland DG. (2006) FoxO are critical mediators of hematopoietic stem cell resistance to physiologic oxidative stress. Cell 2007
