Journal: Cells

Article Title: Reduced Levels of Drp1 Protect against Development of Retinal Vascular Lesions in Diabetic Retinopathy

doi: 10.3390/cells10061379

Figure Lengend Snippet: PCR analysis using mice tail tip DNA indicating genotypes of Drp1 heterozygous knockout ( +/− ) and wild-type (WT) mice. The wild-type Drp1 allele (Drp1 +/+ ) is represented by a band at 0.32 kb, whereas the disrupted allele (Drp1 +/− ) shows a band at 0.54 kb.

Article Snippet: Following blocking, the RTDs were subjected to a primary antibody solution containing mouse monoclonal Drp1 antibody (1:200 in 2% BSA-PBS solution, Catalog #sc-271583, Santa Cruz Biotechnology, Dallas, TX, USA) and incubated overnight at 4 °C in a moist chamber.

Techniques: Knock-Out

Journal: Cells

Article Title: Reduced Levels of Drp1 Protect against Development of Retinal Vascular Lesions in Diabetic Retinopathy

doi: 10.3390/cells10061379

Figure Lengend Snippet: Drp1 +/− mice exhibit reduced Drp1 immunofluorescence in the retinal capillary network. ( A ) Representative images of Drp1 immunofluorescence (green) and DAPI (blue) in retinal capillaries of wild-type (WT), diabetic (DM), Drp1 +/− , and diabetic Drp1 +/− (D-Drp1 +/− ) mice. Scale bar = 100 μm. ( B ) Graphical illustration of cumulative data shows decreased Drp1 immunofluorescence in retinal capillaries of Drp1 +/− mice compared to that of WT mice. * p < 0.01, n = 6; ** p < 0.05, n = 6.

Article Snippet: Following blocking, the RTDs were subjected to a primary antibody solution containing mouse monoclonal Drp1 antibody (1:200 in 2% BSA-PBS solution, Catalog #sc-271583, Santa Cruz Biotechnology, Dallas, TX, USA) and incubated overnight at 4 °C in a moist chamber.

Techniques: Immunofluorescence

Journal: Cells

Article Title: Reduced Levels of Drp1 Protect against Development of Retinal Vascular Lesions in Diabetic Retinopathy

doi: 10.3390/cells10061379

Figure Lengend Snippet: Drp1 expression is normalized in diabetic Drp1 +/− mouse retinas. ( A ) Representative WB image shows Drp1 protein levels in the retinas of WT, diabetic (DM), Drp1 +/− , and diabetic Drp1 +/− (D-Drp1 +/− ) mice. ( B ) Graphical illustration of cumulative data shows diabetes significantly upregulates Drp1 expression and that Drp1 expression is normalized in retinas of D-Drp1 +/− mice. Data are expressed as mean ± SD. * p < 0.01, n = 12.

Article Snippet: Following blocking, the RTDs were subjected to a primary antibody solution containing mouse monoclonal Drp1 antibody (1:200 in 2% BSA-PBS solution, Catalog #sc-271583, Santa Cruz Biotechnology, Dallas, TX, USA) and incubated overnight at 4 °C in a moist chamber.

Techniques: Expressing

Journal: Cells

Article Title: Reduced Levels of Drp1 Protect against Development of Retinal Vascular Lesions in Diabetic Retinopathy

doi: 10.3390/cells10061379

Figure Lengend Snippet: Reduced Drp1 expression lowers Bax activity in diabetic Drp1 +/− mouse retinas. ( A ) Representative WB image shows Bax and cleaved PARP expression in the retinas of WT, diabetic (DM), Drp1 +/− , and diabetic Drp1 +/− (D-Drp1 +/− ) mice. Graphical illustrations of cumulative data suggest reduced Drp1 levels in D-Drp1 +/− mice is protective against diabetes-induced increase in ( B ) Bax levels, ( C ) PARP cleavage and ( D ) caspase-3 activity. Data are expressed as mean ± SD. * p < 0.01, n = 12; ** p < 0.05, n = 12.

Article Snippet: Following blocking, the RTDs were subjected to a primary antibody solution containing mouse monoclonal Drp1 antibody (1:200 in 2% BSA-PBS solution, Catalog #sc-271583, Santa Cruz Biotechnology, Dallas, TX, USA) and incubated overnight at 4 °C in a moist chamber.

Techniques: Expressing, Activity Assay

Journal: Cells

Article Title: Reduced Levels of Drp1 Protect against Development of Retinal Vascular Lesions in Diabetic Retinopathy

doi: 10.3390/cells10061379

Figure Lengend Snippet: Reduced Drp1 level protects against diabetes-induced apoptosis of vascular cells in retinal capillary networks. Representative images of capillary networks showing DAPI-stained cells in the ( A ) WT, ( B ) diabetic (DM), ( C ) Drp1 +/− , and ( D ) diabetic Drp1 +/− (D-Drp1 +/− ) mice. ( E – H ) Corresponding images of TUNEL-positive cells (arrows) in the retinal capillary networks, respectively. ( I – L ) Merged images showing DAPI-stained cells superimposed with TUNEL-positive cells. Scale bar = 100 μm. ( M) Graph of cumulative data showing that retinal capillary networks of diabetic mice exhibited an increase in number of TUNEL-positive cells compared to that of WT mice, while retinal capillary networks of D-Drp1 +/− mice showed reduced number of TUNEL-positive cells compared to that of diabetic mice. Data are presented as mean ± SD. * p < 0.01, n = 6; ** p < 0.05, n = 6.

Article Snippet: Following blocking, the RTDs were subjected to a primary antibody solution containing mouse monoclonal Drp1 antibody (1:200 in 2% BSA-PBS solution, Catalog #sc-271583, Santa Cruz Biotechnology, Dallas, TX, USA) and incubated overnight at 4 °C in a moist chamber.

Techniques: Staining, TUNEL Assay

Journal: Cells

Article Title: Reduced Levels of Drp1 Protect against Development of Retinal Vascular Lesions in Diabetic Retinopathy

doi: 10.3390/cells10061379

Figure Lengend Snippet: Effects of diabetes or decreased Drp1 level on AC and PL development in mouse retinas. ( A – D ) Representative images of retinal trypsin digestion of ( A ) WT, ( B ) DM, ( C ) Drp1 +/− , and D-Drp1 +/− mice show the number of AC (blue arrows) and PL (red arrows) is increased in the diabetic mouse retina compared to that of control mouse retina. Importantly, the number of AC and PL is decreased in the D-Drp1 +/− mouse retina compared to that of diabetic mouse retina. Scale bar = 100 μm. Graphical illustrations of cumulative data show that reduced DRP1 levels in the D-Drp1 +/− mouse retina exhibited a protective effect against the development of ( E ) AC and ( F ) PL. Data are expressed as mean ± SD. * p < 0.01, n = 12.

Article Snippet: Following blocking, the RTDs were subjected to a primary antibody solution containing mouse monoclonal Drp1 antibody (1:200 in 2% BSA-PBS solution, Catalog #sc-271583, Santa Cruz Biotechnology, Dallas, TX, USA) and incubated overnight at 4 °C in a moist chamber.

Techniques: Control