Project description:Purpose: To demonstrate Runx3's association with organizing the extracellular matrix in SFZ chondrocytes. Method: Fragment DNA samples were collected by using Anti-FLAG or IgG, from SFZ chondrocytes treated with lipofection of RUNX3-FLAG Results: 21,730 raw peaks met the peak calling criterion. Conclusions: By GO analysis, the extracellular structure organization and collagen fibril organization terms were the most significantly enriched.
Project description:Purpose: To demonstrate Runx3's association with organizing the extracellular matrix in SFZ/DZ chondrocytes. Method: RNA samples were collected from SFZ/DZ chondrocytes of 5-day-old Col2a1-Cre;Runx3fl/fl and Runx3fl/fl mice. Results: 18 genes were up- or down-regulated by more than 2-fold in the Runx3 knockout in SFZ. Conclusions: However, genes organizing the extracellular matrix were down-regulated in Runx3 cKO DZ chondrocytes.
Project description:To understand difference between SFZ cells and costal chondrocytes, total RNAs from SFZ cells and costal chondrocytes were analyzed by microarray.
Project description:ChIP-seq was conducted using FACS-isolated CD11chiMHCII+CD4+ splenic WT DC and anti-Runx3 antibodies (Ab). Two biological Runx3 IP repeats and two input controls from sorted CD4 DC
Project description:ChIP-seq was conducted using freshly isolated (resting) splenic WT NK cells with anti-Runx3 antibody (Ab), anti-H3K4me1 Ab and non-immune serum (NIS) as control. Runx3 and H3K4me1 IP from splenic NK cells isolated by negative selection using NK cell isolation kit (R&D) followed by sorting of NKp46+ cells.
Project description:ChIP-seq was conducted using freshly isolated (resting) splenic WT CD8+ T cells with anti-Runx3 antibody (Ab), anti-H3K4me1 Ab and non-immune serum (NIS) as control. Two biological Runx3 and two NIS IP repeats from splenic CD8+ T cells isolated by positive selection on anti-CD8 magnetic beads, collected from 18 WT mice.
Project description:Estrogen Receptor alpha (ERα) is a key driver of most breast cancers, and it is the target of endocrine therapies used in the clinic to treat women with ERα positive (ER+) breast cancer. The two methods ChIP-seq (chromatin immunoprecipitation coupled with deep sequencing) and RIME (Rapid Immunoprecipitation of Endogenous Proteins) have greatly improved our understanding of ERα function during breast cancer progression and in response to anti-estrogens. A critical component of both ChIP-seq and RIME protocols is the antibody that is used to pull down the bait protein. To date, most of the ChIP-seq and RIME experiments for the study of ERα have been performed using the sc-543 antibody from Santa Cruz Biotechnology. However, this antibody has been discontinued, thereby severely impacting the study of ERα in normal physiology as well as diseases such as breast cancer and ovarian cancer. Here, we compare the sc-543 antibody with other commercially available antibodies, and we show that 06-935 (EMD Millipore) and ab3575 (Abcam) antibodies can successfully replace the sc-543 antibody for ChIP-seq and RIME experiments.
Project description:ChIP-seq was conducted using splenic WT NK cells cultured for 7 days with IL-2 using anti-Runx3 antibodies (Ab), anti-H3K4me1 Ab and non-immune serum (NIS) as control. Two biological Runx3, one H3K4me1 and two NIS IP repeats from splenic NK cells isolated from individual mice by negative selection using NK cell isolation kit (R&D) cultured with IL-2.