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. Author manuscript; available in PMC: 2018 May 1.
Published in final edited form as: Graefes Arch Clin Exp Ophthalmol. 2017 Jan 25;255(5):955–962. doi: 10.1007/s00417-017-3594-0

Figure 1.

Figure 1

Regulation of oxidative stress prevents glucose-induced decrease in 5mC and increased Dnmt1 binding at the MMP-9 promoter in retinal endothelial cells. Endothelial cells were incubated in 5mM or 20mM glucose for 4 days in the presence or absence of 250μM MnTBAP. (a) 5mC at MMP-9 promoter was quantified using methylated DNA immunoprecipitation kit. (b) Dnmt1 binding was quantified in the crosslinked cells, and IgG was used as a negative antibody control (indicated as ^). Values are represented as mean ± SD from 4–5 samples in each group. (c) Co-localization of Dnmt1 and MMP-9 was performed by immunofluoresence techique using anti-rabbit-DyLight 488 (green) as the secondary antibodies for MMP-9 was anti-mouse-Texas Red (red) for Dnmt1 anti-mouse-Texas Red (red). Cells mounted with DAPI containing medium (blue) w. 5mM and 20mM=cells incubated in 5mM and or 20mM glucose respectively; 20+Mn= cells in 20mM glucose in the presence of MnTBAP; Mann=cells in 20mM mannitol. *p< 0.05 compared to 5mM glucose.