Project description:Transcription profiles of BV2 microglial cell lines: unstimulated, stimulated with LPS or transfected with constitutively active Stat1 and Stat3.
Project description:To comprehensively study the intracellular signaling changes and explore the underlying mechanisms of baicalein-mediated microglial responses, we employed systemic proteomic approach, using target-free SWATH mass spectra (SWATH-MS), focusing on intracellular proteins. BV2 cells were treated with LPS followed by the addition of vehicle or baicalein and they were cultured for a total of 48 hours before being collected for proteomic analysis.
Project description:The OPTN gene is linked to neurodegenerative diseases like amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), which are characterized by chronic microglial activation. Optineurin regulates inflammatory signaling, autophagy, and trafficking, but its role in microglia is not well understood. We used bulk RNA sequencing to profile CRISPR-Cas9-mediated optineurin knockout (KO) and wild-type (WT) BV2 microglia under basal conditions and upon LPS stimulation. At baseline, optineurin KO altered approximately 7% of the transcriptome, with a notable downregulation of type I interferon and antiviral pathways. LPS stimulation in wild-type cells triggered a broad transcriptional shift (~35% of genes). This LPS-induced response was blunted in optineurin-deficient microglia, with ~16% of genes changed relative to the KO baseline. Furthermore, LPS-treated optineurin KO microglia notably diverged from LPS-treated wild-type cells, with ~26% differentially expressed genes. Our findings establish optineurin as a key regulator of the microglial transcriptome. Its loss weakens basal interferon-mediated immune surveillance and decouples the canonical inflammatory response from cell cycle arrest upon activation.
Project description:Mouse microglia (BV2 cells) were stimulated with a TLR4 ligand (E. coli LPS, 1 µg/ml) for 4h. The Agilent SurePrint G3 Mouse Gene Expression Microarray (G4852A) was used for the analysis, which provides full coverage of genes and transcripts with the most up-to-date content, including mRNAs and lincRNAs (http://www.chem.agilent.com/store/en_US/Prod-G4852A/G4852A). BV2 cells were grown to 80% confluence for four groups: the siRNA control (Group A, cells treated with a non-specific scrambied siRNA control), the LPS-stimulated (Group B, cells treated with the siRNA control plus LPS stimulation), lincRNA-Cox2 siRNA (Group C, cells treated with an siRNA to lincRNA-Cox2), and lincRNA-Cox2 siRNA/LPS stimulated (Group D, cells treated with the lincNRA-Cox2 siRNA plus LPS stimulation). Cells were treated with the siRNAs for 24h, followed by additional culture for 4h in the presence or absence of LPS (E. coli LPS, 1 µg/ml). Total RNAs were prepared with the RNeasy Mini kit (Qiagen) according to the manufacturer’s instruction (Ambion).
Project description:This dataset contains the RNA-seq data generated to investigate the mechanism of TOP1 inhibitor topotecan (TPT)'s role of anti-inflammation in microglial. RNA from mouse microgial cells, cells with LPS treatment to induce inflammation, and cells with LPS and TPT treatment were harvest and processed with polyA-tail enrichment RNA library preparation.
