Figure 5.

ApoE was colocalized with MAC on complement-activated RPE cell culture and drusen. (A) RPE cells from a 51-year-old donor with ApoE phenotype E3/E3 and CFH_HH402 variant were primed with S58 (1.2 mg/mL) for 30 minutes and then treated with 6% C1q-Dep for 5 hours. Cells were fixed in 4% PFA for 15 minutes and costained with rabbit anti-human ApoE antibody (green) and mouse anti-human C5b-9 (aE11) antibody (red). Blue stain corresponds to DAPI-stained nuclei. MAC and ApoE were colocalized frequently in the cells. Data are representative of three separate experiments in three donors with similar results. (B) Confocal microscopic colocalization (yellow) of MAC (green) and ApoE (red) in drusen from a 72-year-old donor. Similar results were observed in 94-year-old donor eye (not shown). (C) Primary antibodies were omitted in section of a 72-year-old donor. Scale bar: 50 μm.