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Fig. 6 | BMC Biology

Fig. 6

From: Hyaluronidase-1-mediated glycocalyx impairment underlies endothelial abnormalities in polypoidal choroidal vasculopathy

Fig. 6

Modulating HYAL1 normalizes abnormal cell migration and barrier permeability of PCV endothelial cells. a siRNA knockdown of HYAL1 in BOECs. BOECs were transfected with siRNA for 4 days, including 24 h heterogeneous flow treatment. Cell lysates were harvested and analyzed for HYAL1 levels. Bar graphs show mean actin-normalized HYAL1 band intensities with error bars representing standard deviations (siHYAL1, short-interfering RNA of HYAL1; NT, non-targeting siRNA). b Wound healing assays were carried out to assess migratory capacity of BOECs at 21 h post-scratch after 24 h of static or rotated culture. siRNA knockdown was carried out 48 h prior to orbital rotation. Top panel shows representative image set from the flow condition with yellow outlines indicating cell-free wound regions at 21 h post-scratch, scale bar, 200μm. c Relative KLF2 gene expressions in BOECs in static condition and after 6 h of heterogeneous flow exposure, with treatment of either NT or siHYAL1. d In vitro vascular permeability imaging assay using biotinylated-gelatin-coated surfaces. Intercellular gaps were revealed by Neutravidin-FITC staining after 24 h of static or rotated culture. siRNA knockdown was carried out 48 h prior to orbital rotation. Shown are representative frames from different BOEC lines, scale bar, 500 μm. Area was normalized against average FITC area of normal BOEC lines. All bar graphs showing means with standard deviations. n indicates number of cell lines evaluated for each group while p value is from two-tailed t-test, *p < 0.05

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