School of Science Division of Life Science 16 APOE Variants in Alzheimer's Disease Pathogenesis and Therapy Supervisor: BU Guojun / LIFS Student: WONG Tin Lun / BCB Course: UROP 3200, Summer Alzheimer’s disease (AD) is the leading cause of dementia, with APOE as the most genetic risk determinant: APOE4 significantly increases AD risks, while APOE2 and rare variants (APOE3-Jac and APOE3-Ch) confer protection via different protective mechanisms. Delivery of protective apoE using adeno-associated viral vectors demonstrated a protective effect in an AD mouse model. Based on these findings, a novel, engineered variant, apoE3-Jac-Ch was generated and characterized. This variant was designed to combine the protective biophysical properties of apoE3-Jac (reduces aggregation) and apoE3-Ch (reduces apoEreceptor binding). Preliminary biochemical analyses indicate that apoE3-Jac-Ch reduced both oligomerization and heparin binding affinity comparing to apoE3. These findings suggest that combining these mutations provides additive benefits, highlighting the potential of engineered APOE variants as therapeutic candidates for AD. Molecular Regulation of Skeletal Muscle Stem Cell Quiescence and Activation Supervisor: Tom CHEUNG / LIFS Student: FOO Ashley Sheng Min / BIOT-AB Course: UROP 1100, Spring UROP 2100, Summer This report presents two separate experiments conducted on muscle stem cells (MuSCs). The first experiment investigated the specificity and effectiveness of 5-ethynyl-20-deoxyuridine (EdU) as a cell proliferation marker by comparing EdU incorporation at different time intervals within a 24- and 48-hour culture period. The second experiment evaluated the specificity of two different primary antibodies – Dako and Santa Cruz – used in indirect immunofluorescence staining to detect the myogenic marker MyoD. Both antibodies were applied under identical conditions to assess performance consistency. Together, these experiments highlight the importance of reagent validation in stem cell research, particularly in tracking cell proliferation and ensuring accurate marker detection through immunostaining techniques. Molecular Regulation of Skeletal Muscle Stem Cell Quiescence and Activation Supervisor: Tom CHEUNG / LIFS Student: LUO Wenxin / BCB-IRE Course: UROP 1100, Fall The Click-iT EdU assay is a popular cellular proliferation measurement approach that requires careful control of the overall EdU incorporation ratio to yield effective and valuable results. The aim was to optimize the incubation time for EdU incorporation in two commonly used cell lines, wild-type C2C12 and SH-SY5Y. By examining the percentages of EdU-labeled cells at each incubation interval, specific labeling trends were obtained for each cell line. Both cell lines exhibited a linear trend for EdU incorporation versus incubation time (p < 0.001), which would significantly support a time-dependent labeling kinetics. These results may help with the optimization of incubation times of EdU, enabling experimental designs with more precision when conducting studies on cell replication in these cell lines.
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