Laboratory Intern
Charleston, South Carolina Metropolitan Area
As a part of Medical University of South Carolina's SURP, a NSF-funded REU program, I had the opportunity to receive additional funding from NIH to research in the Gudi lab for 10 weeks to work independently to characterize the role of CPAP, a crucial centrosomal protein, in regulating the endocytic vesicular transport pathway and maintaining EGFR homeostasis. Using a hypomorphic mouse model, I isolated and cultured primary tongue fibroblasts and subjected them to EGF treatment in order to determine the effect of CPAP knockdown on vesicle trafficking. I learned many techniques and procedures to better understand this pathway. Using Western blot analysis, we saw elevated levels of EGFR and thus determined that EGFR was not properly degraded in CPAP knockdown mice, suggesting that the vesicular transport pathway was blocked in mice with reduced CPAP. Confocal microscopy showed significantly less colocalization in the hypomorphic mice, once again suggesting a defect in the pathway. FIJI was used to analyze results. Overall, the endocytic vesicular transport pathway was found to be defective in CPAP-deficient mice.
