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Fig. 3 | BMC Biology

Fig. 3

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

Fig. 3

PCV endothelial cells show attenuated response to heterogeneous flow. a Contour plot overlays on UMAPs of normal and PCV BOECs showing the density distribution of sequenced cells across all clusters. Right panel shows the percentage breakdown of cells per cell state for normal and PCV groups. b Gene set enrichment analysis of GO (Biological processes) on the PCV differentially downregulated genes (in Proinflammatory and Migratory-angiogenic cell states), ranked by Bonferroni corrected p values. c Gene set enrichment analyses of PCV downregulated genes in these clusters were expanded to include Reactome, KEGG, and MSigDB databases. Significantly enriched pathways or processes (adj. p < 0.01) are shown here as violin plots that represent calculated module scores for the gene sets indicated. d (i) Module scores of PCV and normal cells for significantly enriched oxidative stress response and mitochondrial respiration gene sets (adj. p < 0.01) are shown as violin plots here. ii MitoSOX Red staining of PCV and normal BOECs after 24 h of heterogeneous flow. Nuclei were stained DAPI (blue). Right panel shows intensity per cell measurements of MitoSOX Red staining (n > 200 cells analyzed per group, BOECs from 2 PCV and 2 normal individuals). Data shown are normalized to individual static conditions with median and quartiles indicated. ****p < 0.0001 (two-tailed Mann-Whitney test). Scale bar, 100 μm. iii Mitochondrial function assay of BOECs after 24 h of heterogeneous flow. Left panel shows oxygen consumption rate (OCR) of BOECs in response to oligomycin (Oligo.), carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP), and rotenone/ antimycin A (Rot/AA). Right panel shows measurements per well of basal respiration, maximal respiration, and mitochondrial ATP production (BOECs from 4 PCV and 3 normal individuals). Data shown are normalized to individual static conditions with median and quartiles indicated. P values for basal respiration and maximal respiration were from two-tailed t-tests with Welch’s correction and for ATP production from two-tailed Mann-Whitney test, *p < 0.05

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