Fig 1: Effects of EFEMP1 on HUVEC proliferation. CON335 HUVECs, EFEMP1-OE HUVECs, CON077 HUVECs, and shEFEMP1 HUVECs were stained using EdU and Hoechst. EdU-positive cells were highlighted in red and nuclei in blue, following Hoechst staining. Proliferation capacities were analyzed by calculating the percentage of EdU-positive cells. Three independent experiments were performed, and five fields were randomly selected for statistical analysis in each experiment. **p < 0.01.
Fig 2: Effects of EFEMP1 on VEGF expression. VEGF mRNA and protein level alterations in (A) CON335 HUVECs, EFEMP1-OE HUVECs, (B) CON077 HUVECs, and shEFEMP1 HUVECs were detected using qRT-PCR and western blotting, respectively. **p < 0.01.
Fig 3: Effects of EFEMP1 on HUVEC tube formation. (A) Control CON335 HUVECs, EFEMP1-OE HUVECs, (B) control CON077 HUVECs, and shEFEMP1 HUVECs seeded on Matrigel were allowed to form tubes for 12 h in the absence of any additional stimulation. Total tube length, number of branch points, and total number of networks in captured images were measured using MetaMorph. The data represent the average of five independent replicates and the experiments were performed three times. **p < 0.01.
Fig 4: EFEMP1 gene expression in ocular tissues derived from patients with age-related macular degeneration (AMD) and healthy controls from GSE29801. (A) EFEMP1 expression increased in AMD-affected eyes compared with that in the control group. (B) EFEMP1 expression in wet AMD-affected eyes was higher than that in the control group as well as in the pre-AMD and dry AMD groups. **p < 0.01.
Fig 5: (A) EFEMP1 overexpressing (EFEMP1-OE) and (B) EFEMP1 knockdown HUVEC establishment and verification. High-content imaging visualization of HUVECs transfected with GFP-labelled GV492-EFEMP1/GV248-shEFEMP1 vectors was shown. qRT-PCR results of EFEMP1 relative mRNA levels showed a significant increase in GV492-EFEMP1-transfected HUVECs compared with that in GV492-CON335-transfected HUVECs and a significant decrease in GV248-shEFEMP1-transfected HUVECS compared with that in GV492-CON077-transfected HUVECs (**p < 0.01). EFEMP1 protein levels in the four HUVEC groups were tested using western blotting. EFEMP1 protein expression in GV492-EFEMP1-transfected HUVECs increased significantly compared with that in control CON335 and decreased in GV248-shEFEMP1-transfected HUVECS compared with that in control CON077 (**p < 0.01).
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