fmoc nme arg mtr oh  (Chem Impex International)

Journal: Journal of Nanobiotechnology

Article Title: Engineered MEVs for photoreceptor-targeted delivery of USP25 to alleviate diabetic retinopathy

doi: 10.1186/s12951-025-03671-w

Figure Lengend Snippet: MEVs alleviate hyperglycemia-induced photoreceptor injury by upregulating CRYAA expression. ( A ) The proteins in 661 W cells cultured in HG medium with or without MEV treatment were detected using LC-MS/MS. ( B , C ) Representative CRYAA immunostaining images of 661 W cells after MEV treatment and the quantitative analysis of fluorescent intensity compared with the LG group on the basis of three sections per group with three images counted in each section. Scale bars, 25 μm. ( D , E ) Western blot analysis for the CRYAA protein expression in 661 W cells after MEV treatment and the corresponding quantification of protein level compared with the LG group ( n = 3). ( F , G ) Representative retinal CRYAA immunostaining images of db/m and db/db mice after MEV treatment and the quantitative analysis of fluorescent intensity compared with the db/m mice group on the basis of three mice per group with three images counted in each mouse. Scale bars, 100 μm. ( H , I ) Western blot analysis for the retinal CRYAA protein expression in db/m and db/db mice after MEV treatment and the corresponding quantification of protein level compared with the db/m mice group ( n = 3). ( J ) Schematic diagram displaying the experimental design of WT and CRYAA KO mice. ( K ) Western blot to verify CRYAA knockout in the retinal tissues of CRYAA KO mice. ( L ) Fasting blood glucose level testing of WT and CRYAA KO mice at the indicated time points ( n = 6). ( M , N ) Retinal HE staining and ONL thickness measurement in WT and CRYAA KO mice after STZ and MEV treatment ( n = 3). Scale bars, 100 μm. ( O , P ) TUNEL staining of retinas and the analysis of TUNEL + cell percentage of total ONL cells in WT and CRYAA KO mice after STZ and MEV treatment on the basis of three mice per group with three images counted in each mouse. Scale bars, 100 μm. ( Q , R ) Representative retinal rhodopsin immunostaining images of WT and CRYAA KO mice after STZ and MEV treatment and the quantitative analysis of fluorescent intensity compared with the WT + STZ group on the basis of three mice per group with three images counted in each mouse. Scale bars, 100 μm. ( S , T ) Representative retinal s-opsin immunostaining images of WT and CRYAA KO mice after STZ and MEV treatment and the quantitative analysis of fluorescent intensity compared with the WT + STZ group on the basis of three mice per group with three images counted in each mouse. Scale bars, 100 μm. No data were excluded from the analyses. Data collection and analysis were performed by investigators blinded to the experimental groups. n represents independent biological replicates. Data in (C), (E), (G), (I), (L), (N), (P), (R), and (T) are presented as mean ± SEM and were compared using one-way ANOVA with Tukey’s multiple comparison test. ns, not significant, ** P < 0.01, and *** P < 0.001. GCL: ganglion cell layer; INL: inner nuclear layer; ONL: outer nuclear layer

Article Snippet: Wild type (WT) and CRYAA knockout (CRYAA KO ) mice were purchased from Cyagen Bioscience (Suzhou, China) and induced to diabetic model by high-fat diet (HFD) feeding plus injection with streptozotocin (STZ; 30 mg/kg per day; Sigma-Aldrich, MO) intraperitoneally for 7 consecutive days, as in previous studies [ ].

Techniques: Expressing, Cell Culture, Liquid Chromatography with Mass Spectroscopy, Immunostaining, Western Blot, Knock-Out, Staining, TUNEL Assay, Comparison