Project description:Bone marrow-derived MDSCs were generated from bone marrow of naive WT or Rel-/- mice. After red blood cell lysis, BM cells were cultured in complete RPMI medium containing GM-CSF (100 ng/mL) and IL-6 (100 ng/mL) for 7 days. RNAs were collected afterwards for RNA-Seq.
Project description:Fibrosarcoma cell lines were generated from wild-type and Tnfaip8 knockout mice. After stimulation with vehicle, LPA, or PDGF, RNAs were collected from the cells afterwards for RNA-Seq.
Project description:The experiment was designed to determine the gene expression changes cultured brown adipocytes in response to the inflammatory stimulus of LPS treatment. Both wild type and TLR4 knockout cells were applied to enable assessment of the contribution of TLR4 to the response.
Project description:The METTL3 methyltransferase is responsible for the deposition of N6-methyladenosine (m6A) modifications in RNA and has been identified as essential for survival and proliferation of acute myeloid leukemia (AML) cells in a genome-wide CRISPR screen. In our experiments involving a small-molecule METTL3 inhibitor (UZH2) in the AML cell line MOLM-13, we observed suppression of cell proliferation, induction of apoptosis and differentiation. The aim of RNA-seq experiment was to characterize the transcriptomic changes occurring in MOLM-13 cell line after treatment UZH2. Cell were treated with 10 µM of UZH2 for 16 h and compered to untreated controls (5 % DMSO).
Project description:This study is to investigate the potential impact of an lncRNA NR_126553 (Nostril) in murine intestinal epithelial cells (IEC4.1 cells) in response to Cryptosporidium parvum (C. parvum) infection. NR_126553 (Nostril) was knocked down by using a pool of gene specific siRNAs (SiNostril). Cells treated with a scramble non-specific siRNA were used as the control (siNegative control). After siRNA transfection 24h, cells were exposed to C. parvum infection for 24h. Then total RNA was collected for sequencing via BGISEQ-500 platform to obtain a comprehensive view of the transcriptome.
Project description:The experiment looks for diurnal-regulated genes in Medicago truncatula plants. Medicago truncatula Jester accession plants were entrained for18 days in long days (16h light:8h dark) at 24°C constant temperature. Samples for two biological replicates per time point were collected every 4h for 24h beginning at ZT0 (subjective dawn) until ZT20. Total RNA was extracted from ground tissue using an RNeasy RNA extraction kit (Qiagen). Extracted total RNA was DNAse-treated (Invitrogen). Total RNA (2-3ug) was dried down and preserved in Sigma-Aldrich RNAstable before being couriered to BGI Tech Solutions (HONGKONG) Co., Ltd (https://www.bgi.com/global). BGI Tech prepared and processed libraries for 20M PE100 reads using their DNBseq platform - BGISEQ-500.
Project description:The experiment looks for circadian-regulated genes in Medicago truncatula plants. Medicago truncatula Jester accession plants were entrained for 3 weeks in long days (16h light:8h dark) at 24°C constant temperature. On day 20, plants were transferred to LL conditions for 3 days. Samples for three biological replicates per time point were collected every 4h for 24h beginning at CT48 (subjective dawn) until CT68. Total RNA was extracted from ground tissue using an RNeasy RNA extraction kit (Qiagen). Extracted total RNA was DNAse-treated (Invitrogen). Total RNA (2-3ug) was dried down and preserved in Sigma-Aldrich RNAstable before being couriered to BGI Tech Solutions (HONGKONG) Co., Ltd (https://www.bgi.com/global). BGI Tech prepared and processed libraries for 20M PE100 reads using their DNBseq platform - BGISEQ-500.
Project description:RNA sequencing was used to compare the transcriptional response of human infant ileal intestinal enteroids following exposure to human milk exosomes. Exosome-exposed enteroids were compared to enteroids not exposed to exosomes in the same 72 hour period.
Project description:Myeloid-derived suppressor cells (MDSCs) are immature myeloid cells that accumulate in the tumor microenvironment of most cancer patients. There MDSCs suppress both adaptive and innate immune responses, hindering immunotherapies. Moreover, many cancers are accompanied by inflammation, a processes that further intensifies MDSC suppressive activity, causing aggressive tumor progression and metastasis. MDSCs collected from tumor-bearing mice profusely release nano-scale membrane-bound extracellular vesicles, called exosomes, which carry biologically active proteins between cells and contribute directly to the immune suppressive functions of MDSC. Many studies on other cell types have shown that exosomes may also carry microRNAs (miRNAs) and messenger RNAs (mRNAs) which can also be transferred to surrounding and distant cells. However, to the best of our knowledge, the miRNA and mRNA cargo of MDSC-derived exosomes has not yet been interrogated. This study aims to identify and quantify the cargo of MDSC and their immunosuppressive exosomes to gather knowledge that can offer insights on the mechanisms by which MDSCs contribute to immune suppression, focusing on the role of exosomes as intercellular communication mediators in the tumor microenvironment. In order to achieve our objective a well-established mouse model based on a conventional mammary carcinoma (4T1 cells) and heightened inflammation (4T1 transduced to express the cytokine interleukin-1b) was used. We provide evidence that MDSC-derived exosomes carry proteins, mRNAs and miRNAs. Relative quantitation demonstrated quantitative differences between the exosome cargo and the cargo of their parental cells, supporting the hypothesis that selective loading into the exosomes is possible. Additionally, quantitative and functional analyses of the exosome cargo generated under conventional and heightened inflammation conditions are consistent with clinical observations that inflammation is linked to cancer development.