Alterations in ICSBP/IRF-8 and ICSBP/IRF-8 target gene expression and canonical Wnt signaling in Alox15 HSC. (A) Quantitative PCR of mRNA of ICSBP/IRF-8 (▨), ICSBP/IRF-8-regulated genes (■), and genes known to regulate HSC self-renewal but not regulated by ICSBP/IRF-8 (□) in sorted B6 and Alox15 LSK presented as average fold change from B6 (n = 3). (B-C) Decreased canonical Wnt signaling in Alox15 HSC. (B) Immunoblot (top) and densitometric quantification (bottom) of β-catenin, a downstream mediator of canonical Wnt signaling, in lysates of B6 and Alox15 Lin⁻cKit⁺ cells are shown (n = 4). (C) Decreased percentage of Alox15 LSK expressing β-catenin in the nucleus determined by immunofluorescence of sorted cells. Representative images of B6 and Alox15 LSK β-catenin or isotype-matched control (FITC) staining and DAPI (Blue) are shown at an original magnification ×40, with a summary of results; *P < .05, **P < .01 compared with wild-type. All error bars represent the average (± SEM).