Project description:SPARC-deficient mice have been shown to exhibit impaired glucose tolerance and insulin secretion, but the underlying mechanism remains unknown. We used microarrays to detail the global programme of gene expression and identified distinct classes of up-regulated and down-regulated genes changed by overexpressing SPARC.
Project description:To further explore the underlying molecular mechanism of GR’s effects, we transfected the Min6 cell with the GR cDNA expression vector or control vector, and then extracted RNA of Min6 cells. M6A antibody was used to specifically recognize m6A on mRNA. methylation modified RNA fragments were obtained by immunoprecipitation and used for high-throughput sequencing. By comparing the captured RNA fragments, we found that there were 22381 difference peaks between GR overexpression group and control group, among which 3659 binding peaks were significantly different.Further analysis the MeRIP sequencing results of MIN6 cells overexpressing GR revealed that decreased m6A was detected in a large number of autophagy-related mRNAs.
Project description:The microRNAs miR-141 and -200c were co-overexpressed in the Min6 beta-cell line, which caused pro-apoptotic gene expression and subsquently apoptosis. We used Affymetrix Chip analysis to address which genes are regulated upon miR-200-overexpression to define target genes of miR-200 and pro-apoptotic genes upregulated by miR-200 to induce apoptosis. Min6 cells were infected with a control Adenovirus or Adenovirus overexpression miR-141/200c. Cells were harvested with Trizol and RNA extraction was performed. RNA from n=3 wells was pooled for each condition and subjected to Affymetrix chip analysis
Project description:Comparative analysis of the transcriptome of 4T1 cells stably transduced with a lentiviral vector expressing a siRNA against murine SPARC (4T1-C18) with 4T1 control cells stably transduced with a lentiviral vector expressing a scramble sequence (4T1-SCR). Two-condition experiment, 4T1-C18 vs. 4T1-SCR cells. Biological replicates: 4 SPARC knock down, 4 control, independently grown in vitro and harvested. One replicate per array. Microarrays were hybridized in three different days.
Project description:To determine the impact of Hnf1a overexpression on gene expression in mouse beta cells, we performed RNA-seq experiments on MIN6 where Hnf1a was overexpressed using CRISPR activation (SAM system).
Project description:The cells and mechanisms involved in blood clot resorption are only partially known. Regulatory T cells (Treg) accumulate in venous blood clots and regulate thrombolysis by controlling the recruitment, differentiation and matrix metalloproteinase (MMP) activity of monocytes. The clot Treg population is heterogeneous and contains a population of Treg that forms the matricellular acid- and cysteine-rich protein (SPARC). SPARC induces MMP activity in monocytes and SPARC+ Treg are required for clot resorption.
Project description:The aim of this study was evaluate the transcriptome changes in the comparison between triple negative tumors with increased SPARC expression and triple negative tumors with decreased SPARC expression according to Nagai et al., 2011 (Breast Cancer Res Treat (2011) 126:1–14) The results generated could be of particular interest to better define the prognostic impact of SPARC expression in triple negative breast tumors