Project description:The goal of this experiment is to determine the expression levels of the genes that are influenced by the downregulation of plek2. TER119 negative mouse fetal liver erythroblasts were purified and infected with control and plek2 shRNA retroviruses and cultured in Epo free medium for 24 hours followed by Epo medium for 24 hours. Microarray gene expression profile analysis was performed using an Illumina platform with a biological triplicate of the same experiment.
Project description:We identified genes expressed in mouse liver that are regulated by Cux2, a highly female-specific liver transcription factor whose expression is regulated by sex-dependent plasma GH patterns. Using siRNA to knockdown Cux2 expression in female liver, we show that female specific genes are predominantly repressed by Cux2 knockdown. In contrast, similar numbers of male-biased genes are repressed as are induced by Cux2 knockdown. A scrambled, non-specific siRNA was used as a control. (Published in: TL Conforto et al 2012, Mol Cell Biol. 2012, 32:4611-4627. PubMed PMID: 22966202; PMCID: PMC3486175)
Project description:The fetal hemoglobin (HbF) levels were 95.4 ± 1.5% and 4.4 ± 0.2% in fetal liver-derived and adult blood derived-cultured erythrocytes (n=5), respectively. Following RNA isolation from the CD71 high/GPA positive erythroblasts, gene expression analyses were performed using Affymetrix Human Gene 2.0 ST Array. The Affymetrix raw data files of microarray were preprocessed using robust multi-array average method for background correction, log-transformation, and quantile normalization. Differential gene expression between fetal liver derived- and adult peripheral blood derived-erythroblasts was determined using moderated t-statistics implemented in the Limma package. We found the 1366 genes were upregulated by more than 1.5-fold change with adjusted p-value less than 0.05 in fetal liver derived-erythroblasts.
Project description:V617F driver mutation of JAK2 is the leading cause of the Philadelphia-chromosome-negative myeloproliferative neoplasms (MPNs). Loss of Plek2 ameliorated JAK2V617F-induced myeloproliferative phenotypes including erythrocytosis, neutrophilia, thrombocytosis, and splenomegaly, thereby reverting the widespread vascular occlusions and lethality of JAK2V617F knockin mice. To reveal the role of Plek2 in the pathogenesis of JAK2V617F-induced MPNs and the detail mechanisms of its rescue, we performed RNA sequencing to analyze the gene expression profiles change between JAK2V617F/+ Plek2+/+ and JAK2V617F/+ Plek2-/- erythroblasts and hematopoietic stem/progenitor cells.
