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. 2021 Jul 13;108(8):1367–1384. doi: 10.1016/j.ajhg.2021.06.002

Figure 5.

Figure 5

Immunohistochemistry of human retinas shows localization of FHR-2 and FHR-5 in the choriocapillaris and in drusen

GC = ganglion cells; BC = bipolar cells; PR = photoreceptors; OLM = outer limiting membrane; RPE = retinal pigment epithelium; CC = choriocapillaris, and BM = Bruch’s membrane.

To circumvent the difficulties originating from the intense autofluorescence of retinal/choroidal structures (e.g., lipofuscin granules, drusen, and capillary walls), we used a non-fluorescent immunolabeling method to detect FHR-2 and FHR-5 in a non-AMD eye. The distributions of the two proteins are indistinguishable and confined to the choroid (A and C, for FHR-2 and FHR-5, respectively; the scale bar represents 100 μm). The highest signal intensities were detected in the connective tissue surrounding the choriocapillaris. Similarly, strong signal can be detected in the Bruch’s membrane and in drusen (B and D, for FHR-2 and FHR-5, respectively; the scale bar represents 20 μm). Sections were incubated with the primary antibodies anti-FHR-2.1 from Sanquin for FHR-2 and anti-FHR-5.4 from Sanquin for FHR-5. The antibody anti-FHR-2.1 was generated against FHR-2 but also recognizes FHR-1. Anti-FHR-5.4 is monospecific for FHR-5; therefore, staining with a monoclonal antibody found to recognize FHR-2 exclusively (R&D Systems, MAB5484) was also performed (Figure S5).