Figure 10.

Repetitive complement challenge enhanced cell-associated ApoE accumulation. RPE cells from a 62-year-old donor with ApoE phenotype E3/E3 and CFH_YY402 variant were primed with or without S58 (1.2 mg/mL) for 30 minutes and then treated with either 6% C1q-Dep or HiC1q-Dep for 24 or 48 hours. For the second and third complement challenges, cells were washed once, reprimed with or without S58, and then incubated with 6% C1q-Dep or HiC1q-Dep for 48 hours. Repetitive challenge was performed every other day for 1 week (three challenges). Total proteins (15 μg) were separated by SDS-PAGE. The relative quantity of ApoE protein normalized to GAPDH is shown below each lane. Blot images are representative of seven separate experiments in three donors with similar results. The bar graph depicts pooled data from two separate experiments in which cells from a single donor were treated under identical conditions.