Fig. 2

Abnormal DNA Hypermethylation of Pltp Promoter is Associated with DR. A Venn diagram of the DMG and DEG. B Expression of three key genes (Pltp, Prrt1, and Aen) with abnormal DNA methylation in the retina (NC vs DR mice, n = 3). C Agarose gel electrophoresis results of MS-PCR products of the key genes and quantitative analysis of promoter methylation levels (NC vs DR mice, n = 5). D Western blot analysis of PLTP in retina (NC vs DR mice, n = 3). E Single-cell sequencing showing Pltp predominantly distributed in retinal cells ((NC vs DR mice, n = 3). F Retinas from mice with diabetic retinopathy and non-diabetic controls were incubated with antibodies against PLTP and vWF, followed by secondary antibodies conjugated with Alexa Fluor 488 and Alexa Fluor 647 respectively (NC vs DR mice, n = 3). Red boxes show areas that co-express vWF and PLTP which surround retinal endothelial cells. DMG, differentially methylated genes; DEG, differentially expressed genes; MS-PCR, methylation-specific PCR; M, methylation bands; U, unmethylated bands; M/U, quantifying the ratio of the intensity of the methylated band to the unmethylated band; NC, normal control; DR, diabetic retinopathy; vWF, von Willebrand factor. *p < 0.05, **p < 0.01, ***p < 0.001