RNA sequencing analysis of human embryonic stem cell line BG01 treated with anti-MFG-e8 neutralizing antibodies (anti-M8nAb)
To understand the function of MFG-e8, which is secreted from human embryonic stem cells (hESCs), hESC line BG01 cells were treated with anti-M8nAb to deplete the MFG-e8 activity. Depletion of MFG-e8 activity in hESC cultures resulted in decreased cell proliferation, increased cell death, and prominent morphological changes within 2 days. To investigate the transcriptomic changes under depletion of MFG-e8 activity in hESC culture, we performed RNA-seq analysis of BG01 cells treated with anti-M8nAb at two different time point. As a result, we identified differentially expressed genes in anti-M8nAb treated BG01 cells compared to isotype IgG treated control cells. Transcriptome of anti-M8nAb tretaed BG01 cells were enriched with the genesets related with cell death, cell cycle arrest, and epithelial-to-mesenchymal transition compared to isotype IgG treated control cells. In conclusion, these RNA-seq analysis results can provide insights into how MFG-e8 affect signaling mechanisms in hESCs. Overall design: To investigate the function of MFG-e8 in maintaining hESC culture, we treated hESC line BG01 with 5 µg/ml anti-M8nAb to deplete MFG-e8 activity. Normal mouse IgG (5 µg/ml) was treated as isotype control of anti-M8nAb. (2 biological replications for each samples) We performed gene expression profilling analysis using RNA-seq data of the BG01 cells treated with anti-M8nAb or isotype IgG for 12 or 24 hours.