Project description:To investigate the function of lymphotoxin alpha in the regulation osteoblasts, murine pre-osteoblast cell line MC3T3-E1 clone #4 was treated with recombinant human lymphotoxin alpha1/beta2 for different times. Subsequently, we extracted total RNA from the treated cells, performed reverse transcription and cDNA library for RNA-Seq analysis.
Project description:To explore the mechanism by which osteoblastic 11β-HSD1 regulates bone formation and glucose handling, we transfected the mouse MC3T3-E1 osteoblastic cells with a plasmid carrying the exogenous Hsd11b1 or EGFP gene to construct the 11β-HSD1-overexpressed and control osteoblastic cells (hereafter MC3T3-HSD1 and MC3T3-GFP cells), respectively. After incubating with 11-DHC,we then employed RNA sequencing (RNA-seq) to examine the gene expression profile between MC3T3-HSD1 and MC3T3-GFP cells.
Project description:Purpose: This is a post-GWAS functional study that aims to determine the effect of Akap11 knockout in mouse preosteoblastic cell line MC3T3-E1 during osteogenesis. Methods: mRNA profile of WT (treated with CRISPR and remains unedited) and Akap11 knockout MC3T3-E1 cells were generated by RNAseq from pooled RNA (from 3 biological triplicate at equal quantity) of various timepoint during osteogenesis (Day 0, 7,16, 25).100ng total RNA was used for library construction by KAPA Stranded mRNA-Seq Kit (Roche). Pair-End 101bp sequencing was performed in the Illumin HiSq1500 sequencer using the HiSeq SBS Kit v4 (Illumina). Data was collected with SCS version 1.4.8. Base calling was performed using Illumina’s RTA (Version 1.12.4.2). 7 samples were sequenced per lane and achieved a sequencing depth of ~65 million read per sample. Results: Expression of osteogentic markers in Akap11 knockout MC3T3-E1 cells were suppressed. Genes invovled in IGF-I signaling pathway were signficantly altered.
Project description:To explore the mechanism by which osteoblastic 11β-HSD1 regulates bone formation and glucose handling, we transfected the mouse MC3T3-E1 osteoblastic cells with a plasmid carrying the exogenous Hsd11b1 or EGFP gene to construct the 11β-HSD1-overexpressed and control osteoblastic cells (hereafter MC3T3-HSD1 and MC3T3-GFP cells), respectively. After incubating with 11-DHC,we then employed assay for transposase-accessible chromatin with sequencing (ATAC-seq) to examine the gene expression profile between MC3T3-HSD1 and MC3T3-GFP cells.
Project description:In order to define the underlying mechanism of fluoride resistance in mammals and provide a theoretical basis for fluorosis treatment, high-throughput sequencing was applied to map the genetic changes of fluoride-resistant mouse osteoblasts. Fluoride-tolerant MC3T3-E1 cells were developed by gradient fluoride exposure. The differentially expressed genes of fluorine-resistant MC3T3-E1 cells were identified by high-throughput sequencing. High-throughput RNA sequencing identified 2702 differentially expressed genes (DEGs) showed more than 2-fold difference in 30ppm FR MC3T3-E1 cells, of which 17 DEGs were associated with ferroptosis.
Project description:In this study, we compared the gene expression profiles of mouse MC3T3-E1 cells treated with PAKG MPs to those of the control group.
Project description:Transcriptional profiling of MC3T3-E1 osteoblasts that were flow cytometry-separated from cocultures with control or Jagged1-overexpressing tumor cells and treated with either DMSO control or 1μM MRK-003 (gamma-secretase inhibitor). One cell line (MC3T3-E1) cells: four different experimental conditions: cultured with (1) control tumor cells + DMSO; (2) Jagged1-overexpressing tumor cells + DMSO; (3) control tumor cells + MRK-003; (4) Jagged1-overexpressing tumor cells + MRK-003. Each experiment has two biological replicates. Total, 8 samples.
Project description:Bone is the main site of metastasis from prostate cancer, it is important to investigated miRNAs and mRNAs of bone metastases from prostate cancer. Considering that bone is in an appropriate mechanical environment in physiological state, in this study, the miRNA, mRNA, lncRNA profiles of mechanically strained osteoblasts treated with conditioned medium of PC-3 prostate cancer cells were studied. MC3T3-E1 osteoblastic cells were treated with conditioned medium of PC-3 prostate cancer cells, at the same time stimulated with mechanical tensile strain of 2,500 microstrain (με) at 0.5 Hz, the osteoblastic differentiation of the MC3T3-E1 cells were assayed
