Fig. 7

Short-term co-culture of stretched HUVECs induces ARPE-19 cytotoxicity and TJ disruption. A Cell viability of ARPE-19 detected by CCK-8. ARPE-19 cells are treated with conditioned medium from unstretched (Co-ctr) or stretched (Co-Stretch) HUVECs for 24 h. The blank group refers to ARPE-19 cells that have not been co-cultured (n = 5). CCK-8, Cell Counting Kit-8. B Images of BrdU–labeled ARPE-19 cells co-cultured for 24 h. BrdU, 5-iodine-2'-deoxyuridine. C Percentage of the shown BrdU+ cells (n = 5). D Images of scratch wound healing assay for ARPE-19 cells co-cultured for 0, 12, 24, 36, 48 or 60 h. E Quantification of unhealing wound area compared to the baseline (0 h, n = 4). F Expression of TJ proteins ZO1 and Occludin in ARPE-19 cells co-cultured for 24 h suggested by immunoblotting. GAPDH served as the internal control. TJ, tight junction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. G Quantification of shown blots, normalized by GAPDH (n = 3). H Immunofluorescence staining of TJ marker ZO1 and Occludin in ARPE-19 cells co-cultured for 24 h. Data in (A, C, E, G) are presented as mean ± SD. *p < 0.05, and ***p < 0.001