🧫 Experiment Protocol
ExploratoryAtherosclerosisC1QA, C1QC, SPI1RAW264.7 macrophagesproposed
This cell culture experiment was designed to validate the expression of identified hub genes in an in vitro atherosclerosis model. RAW264.7 macrophages were treated with oxidized low-density lipoprotein (ox-LDL) to simulate the atherosclerotic environment. The expression levels of C1QA, C1QC, and the transcription factor SPI1 were measured using quantitative real-time PCR (qRT-PCR). This experiment aimed to confirm that the identified hub genes are indeed upregulated in macrophages exposed to atherogenic stimuli, thereby validating the computational findings in a controlled cellular system that mimics key aspects of atherosclerotic plaque formation.
PRIMARY OUTCOME
Validation of high expression levels of C1QA, C1QC, and SPI1
EXPECTED OUTCOMES
Increased expression of C1QA, C1QC, and SPI1 in ox-LDL treated macrophages
SUCCESS CRITERIA
Confirmation of upregulated gene expression in treated versus control cells
PROTOCOL
Treatment of RAW264.7 macrophages with ox-LDL followed by qRT-PCR analysis of gene expression