Project description:The aim of the study was to characterize the role of PCSK9 in human beta cells. We performed siRNA-mediated knockdown of PCSK9 in human beta cell line EndoC-bH1 and compared the expression profiles against control siRNA-treated cells.
Project description:KCTD17 is downregulated in human T2D beta-cells. To investigated the impact of KCTD17 ablation in beta-cells we performed loss of function experiments in EndoC-bH1 cells.
Project description:N6-methyladenosine (m6A) is the most abundant chemical modification in mRNA. However, the role of m6A and the precise mechanisms involved in the beta-cell adaptation to metabolic demand are unexplored. Here, we performed enhanced crosslinking and immunoprecipitation (eCLIP) assays to identify m6A sites regulated by ALKBH5 in EndoC-bh1 human beta-cells.
Project description:EndoC-BH1 cells were plated in normal media and forward transfected with lipofectamine 3000 (2ug cDNA) of empty, OAS1, OAS2, or OAS3 plasmids from Vector builder
Project description:We have developped a novel human pancreatic beta cell line: EndoC-βH5. EndoC-βH5 cells are ready-to-use and storable cells with physiological insulin secretion. EndoC-βH5 cells were generated by integrative gene transfer of immortalizing transgenes hTERT and SV40 large T along with Herpes Simplex Virus-1 thymidine kinase into human fetal pancreas. Immortalizing transgenes were removed after amplification using CRE activation and remaining non-excized cells eliminated using ganciclovir. Resulting cells were distributed as ready to use EndoC-βH5 cells. We performed comparative transcriptome analysis with EndoC-βH1 cells , extensive functional and immunological assays. RNA-seq confirmed abundant expression of beta cell transcription factors and functional markers, including incretin receptors.
Project description:RFX6 is a key transcription factor for the development of mouse pancreas, however the functional roles of RFX6 in human beta cells are poorly explored. Thus transcriptome analysis was perfomed to determine the functional targets of RFX6 in human beta cells using the recently developed human beta cell-line EndoC-M-NM-2H2. Transcriptome profile of human beta cell line (EndoC-M-NM-2H2 cells) following siRNA induced knockdown of RFX6 is compared with siControl (siNT) treated EndoC-M-NM-2H2 cells.
Project description:EndoC-bH1 cells were incubated with control siRNA or PCSK9 siRNA for 72 hours. The experiment was powered based PCSK9 knockdown in a pilot experiment. Sample size: 12x12.