Fig. 1: ManN is a mitogen/survival factor for eye-derived microvascular endothelial cells. | Nature Communications

Fig. 1: ManN is a mitogen/survival factor for eye-derived microvascular endothelial cells.

From: Inhibition of protein glycosylation is a novel pro-angiogenic strategy that acts via activation of stress pathways

Fig. 1

a Bell-shaped effects of Mannosamine (ManN) on bovine choroidal microvascular endothelial cells (BCEC) proliferation. BCECs were treated with various concentrations of ManN ranging from 0.5 μM to 1 mM for 5–6 days, with or without 5 ng/ml VEGF. At the end of the experiment, cells were fixed and stained with crystal violet. Cell-covered areas in various treatment groups were quantified by ImageJ software. b Cell numbers were quantified by AlamarBlue® assay and fluorescence was measured at 530 nm/590 nm. n = 3 independent samples. c Effect of ManN on bovine retinal microvascular endothelial cells (BREC) proliferation. n = 3 biologically independent samples. d Effects of hexosamines other than ManN on BCEC proliferation. Each treatment group was tested in duplicate. e BCEC confluent monolayers were scratched with 1 ml pipet tip, washed and then incubated for 40 h in low-glucose DMEM containing 1% FBS. n = 3 independent samples. Scale bar = 400 µm. Images were taken and gaps between leading wound front were quantified using AxioVision LE Rel.4.4 software. Representative images from crystal violet staining are shown. f Effects of ManN in BCEC transwell migration assay. n = 4 independent samples. Asterisks indicate a significant difference compared with control. When statistical analysis was done using a different control, a line was used between specific groups. A representative experiment is shown in two independent studies. Data are means +/− SD, Statistical analysis was done by two-tailed, two-sample unequal variance t test. *p < 0.05, **p < 0.01. Data are provided as a Source data file.

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