Project description:GAS2DN could suppress the growth of chronic myeloid leukemia cells, including K562, MEG-01 and CD34+ cells from patients. In addition, GAS2DN inhibited the tumorigenic ability of MEG-01 cells in nude mice. To understand the molecular insight of this inhibitory effect of GAS2DN, global gene expression were performed. The control and GAS2DN-transduced MEG-01 cells were used for microarray analysis.
Project description:GAS2DN could suppress the growth of chronic myeloid leukemia cells, including K562, MEG-01 and CD34+ cells from patients. In addition, GAS2DN inhibited the tumorigenic ability of MEG-01 cells in nude mice. To understand the molecular insight of this inhibitory effect of GAS2DN, global gene expression were performed. The control and GAS2DN-transduced MEG-01 cells were used for microarray analysis. Three biological independent extracts of control and GAS2DN-transduced cells were pooled together with equal amount, and then the pooled samples were compared with Affymetrix chips.
Project description:Proteomic analysis of cell-free supernatants (Secretome) from human platelets (n = 4, donors) and megakariocytes (MEG-01, n = 3 replicates).
Project description:SON is a large Ser/Arg (SR)-related protein localized in nuclear speckles. SON siRNA causes defects in mitotic progression and genome instability. We used microarrays to detail the pattern of gene expression after SON knockdown.
Project description:SON is a large Ser/Arg (SR)-related protein localized in nuclear speckles. SON siRNA causes defects in mitotic progression and genome instability. We used microarrays to detail the pattern of gene expression after SON knockdown. HeLa cells were grown in 100 mm dishes and transfected with negative control siRNA or SON siRNA (400 pmol). Then, cells were harvested after 66 hours and RNAs were isolated. Microarrays were performed using GeneChip Human Genome U133A 2.0 (Affymetrix).
Project description:Exon and expression analysis of HeLa cells after knockdown of SON Serine-arginine-rich (SR) proteins play a key role in alternative pre-mRNA splicing in eukaryotes. Our laboratory recently showed that a large SR protein called Son has unique repeat motifs that are essential for maintaining the subnuclear organization of pre-mRNA processing factors in nuclear speckles. Motif analysis of Son highlights putative RNA interaction domains that suggest a direct role for Son in pre-mRNA splicing. A genome-wide screen was performed to identify putative human transcription and splicing targets of Son. HeLa cells were transfected with siRNA against SON or a control siRNA (siLuciferase) for 48 hours. Five biological replicates were used for each condition.
Project description:Transcriptional profiling of the effect of shRNA silencing of Runx1 in human AMkL Meg-01 cells. RNA samples obtained from two independent colonies were compared to RNAs from negative control transductions in a two-color design. A total of four microarrays were completed, with a dye-swapped pair performed for each colony.
Project description:Transcription factor GFI1B suppresses its own gene expression, and deregulated GFI1B expression contributes to myeloid neoplasms. The DNA polymorphism rs621940 located 8 kb downstream of GFI1B predisposes for clonal hematopoiesis and myeloproliferative neoplasms. Whether rs621940 affects GFI1B expression is unknown. We observed clear GFI1B binding around the rs621940 locus in megakaryoblast MEG-01 cells using ChIP-seq. Pull-downs using DNA-sequences with either rs621940-allele followed by protein identification using mass spectrometry detected 24 differentially bound proteins in MEG-01 cells, but GFI1B was not among these. Also, GFI1B auto-repression was not different between either rs621940-allele in transient gene reporter assays. Deletion of an 1.6 kb fragment encompassing rs621940 in K562 cells did not alter GFI1B expression nor GFI1B auto-repression. In primary peripheral blood mononuclear cells from healthy donors, absolute GFI1B expression did not differ consistently between the rs621940 alleles. Finally, GFI1B haplotyping using long range DNA sequencing detected 3’ non-coding polymorphisms on the same chromosome as the rs621940 polymorphism. Yet, targeted deep RNA sequencing did not detect consistent effects of rs621940 on allelic GFI1B expression. These studies find no evidence that rs621940 affects GFI1B expression nor GFI1B auto-repression.
Project description:Transcriptional profiling of the effect of shRNA silencing of Runx1 in human AMkL Meg-01 cells. RNA samples obtained from two independent colonies were compared to RNAs from negative control transductions in a two-color design. A total of four microarrays were completed, with a dye-swapped pair performed for each colony. Two-condition experiment, Runx1 knockdown vs. negative transduction controls. Biological replicates: 2 knockdown replicates, 2 control replicates.
Project description:chIP-on-chip analysis of GATA1 in the megakaryoblastic cell line Meg01. The IP was obtained with a GATA1 C-terminus antibody. The control sample was a normal goat IgG pull down done side by side with the GATA1 (C-20) antibody using the same soluble chromatin sample from non-treated Meg-01 cells. A 2-chip microarray set (Agilent 240K) was used for the chIP-on-chip analysis.