Project description:We found that a novel gene Gm614 mRNA significantly changed in plasma cells. Because of plasmablast-like, Mus musculus myeloma SP 2/0 cell line was selected to test the effect of Gm614 overexpression on plasmablast/plasma cells. Gm614 cDNA (General Biosystems, Anhui, China) was cloned into lentiviral vector LV201 (Fugene Corp., Guangzhou, China) to generate Gm614 protein. Gm614-expressing LV201 was then transfected into SP 2/0 cells, and stable transfectants were identified by drug selection (Puromycin, Sigma, 10 μg/ml). To identify the effect of Gm614 overexpression on gene expression, we determined mRNA profiles in Gm614-overexpressed and vector-transduced SP 2/0 cells by RNA-seq. RNA-seq was done with an Illumina HiSeq 2500 instrument at GENEWIZ, Suzhou, China.
Project description:To investigate the impact of Stxbp6 knockout on the brain, we carried out transcriptome analysis on the cortexes of Stxbp6-null (n = 3, KO) and wildtype mice (n = 3, WT) and identified 126 differentially expressed genes (DEGs) in the KO group, of which 57 were upregulated and 69 were downregulated. RNA-seq was performed on the BGISEQ platform by MGI Technology Co., Ltd, Guangdong, China.
Project description:Total RNA was extracted from WT and RBP-JCKO mBMDMs at 36 hpi (HTMV, MOI=1), using TRIzol (Invitrogen). Ribosomal RNA was removed using the Ribo-Zero™ kit (Epicentre Biotechnologies). Fragmented RNA (the average length was approximately 200 bp) was subjected to first-strand and second-strand cDNA synthesis followed by adaptor ligation and enrichment with a low cycle according to the instructions of the NEBNext® Ultra™ RNA Library Prep Kit for Illumina (NEB). The purified library products were evaluated using the Agilent 2200 TapeStation and Qubit®2.0 (Life Technologies). The libraries were paired-end sequenced (PE150, sequencing reads were 150 bp) at Guangzhou Ribo Biotechnology (Guangzhou, China) using the Illumina HiSeq 3000 platform. Transgenic mice type: RBP-JCKO (Lyz2-Cre+ RBP-Jfloxed) C57BL/6J Mice.
Project description:We found that a novel gene LOC108168067 mRNA significantly changed in plasma cells. Because of plasmablast-like, mus musculus myeloma SP 2/0 cell line (ATCC® CRL-1581™, https://www.atcc.org/products/all/CRL-1581.aspx) was selected to test the effect of LOC108168067 overexpression on plasmablast/plasma cells. LOC108168067 cDNA (General Biosystems, Anhui, China) was cloned into lentiviral vector LV201 (Fugene Corp., Guangzhou, China) to generate LOC108168067 protein. LOC108168067-expressing LV201 was then transfected into SP 2/0 cells, and stable transfectants were identified by drug selection (Puromycin, Sigma, 10 μg/ml). To explore the effect of on gene expression, we determined mRNA profiles in LOC108168067-overexpressed and vector-transduced SP 2/0 cells by RNA-seq. RNA-seq was done with an Illumina HiSeq 2500 instrument at GENEWIZ, Suzhou, China.
Project description:Total RNAs from exosomes was used for miRNA library preparation and sequencing.The preparation and sequencing of exosome miRNA were performed by Ribobio (Guangzhou, China). First, total RNA samples were fractionated by using 15% Tris-borate-EDTA (TBE) polyacrylamide gel (Invitrogen), then small RNAs ranging from 18 to 30 nucleotides were used for library preparation. Amplification of these small RNAs was then accomplished by PCR. Next, the Illumina HiSeq 2500 platform was used to sequence these RNAs.
Project description:In order to study the role of the HDAC9 in human mesenchymal stem cell differentiation, gene expression analysis was performed with inducible silencing of HDAC9 in human mesenchymal stem cell purchasing from Cyagen Biosciences (HUXMA-90011, Guangzhou, China). Transcriptomic analysis performed on mRNA of human mesenchymal stem cells transfected with lentiviral knockdown HDAC9 (lenti-HDAC9) particles revealed down and up regulation of transcripts of hMSCs differentiation genes
Project description:We found that a novel gene BC094916 mRNA significantly decreased in plasma cells. Because of plasmablast-like, mus musculus myeloma SP 2/0 cell line was selected to test the effect of BC094916 overexpression on plasmablast/plasma cells. BC094916 cDNA (General Biosystems, Anhui, China) was cloned into lentiviral vector LV122 or LV201 (Fugene Corp., Guangzhou, China) to generate BC094916 and BC094916-EGFP fusion protein, respectively. BC094916-expressing LV122 was then transfected into SP 2/0 cells, and stable transfectants were identified by drug selection (Puromycin, Sigma, 10 μg/ml). BC094916 overexpression suppressed SP 2/0 cell proliferation by inducing cell apoptosis. Importantly, BC094916 overexpression effectively suppressed tumor progression in the SP 2/0 xenograft mouse model. In addition, we found that BC094916 is a suppressive transcriptional factor. To verify its target genes, we determined mRNA profiles in BC094916-overexpressed and vector-transduced SP 2/0 cells by RNA-seq. RNA-seq was done with an Illumina HiSeq 2500 instrument at GENEWIZ, Suzhou, China.
