Figure titles and legends
Figure 1A. Family pedigrees with juvenile-onset open-angle glaucoma (JOAG) and EFEMP1 mutation status. Arrow indicates the proband. Squares indicate male subjects; and circles refer to female subjects. Slash through indicates a deceased individual. Solid symbols indicate subjects diagnosed with JOAG; half-filled symbols indicate subjects who carry the EFEMP1 variant but with no clinical signs of glaucoma; and open symbols refer to subjects with no glaucoma and noEFEMP1 variant. Genotypes are heterozygous mutant (m/+) and wild type (+/+). Figure 1B. Schematic diagram of EFEMP1 and evolutionary conservation. EFEMP1 Genbank transcript NM_001039348.3 and color-coded exons corresponding to the protein domain. The mutations p.Asn80Tyr, p.Arg477Cys, and p.Ter494Glnext*29 are located at highly conserved domains across several organisms.
Figure 2. Fundus images from a patient from Family B with EFEMP1 variant p.Ter494Glnext*29 (B-V:5). The image show damage to the optic nerve but no evidence of subretinal deposits (drusen) characteristic of Malattia Leventinese/Doyne Honeycomb dystrophy.
Figure 3. Single cell RNA sequencing EFEMP1 expression and known early onset glaucoma genes from the human aqueous humor outflow pathway. Comparative expression levels of EFEMP1 and genes known to cause various types of early onset glaucoma. Plot generated from the Broad Institute of MIT and Harvard’s Single Cell Portal using data from “Cell atlas of aqueous humor outflow pathways in eyes of humans and four model species provides insight into glaucoma pathogenesis” by van Zyl et al, 2019.16
Figure 4 . Expression of EFEMP1 in COS-7 cells.Cultured COS-7 cells transfected with wildtype and variant EFEMP1 and processed with immunocytochemistry and the following stains: DAPI (nucleus), CellLight ER-RFP (endoplasmic reticulum), Alexa 647 (EFEMP1protein), GFP (GFP expression reporter). Imaged using Leica SP8 confocal microscope using 63x objective (glycerol immersion), digital zoom 2x.