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mdivi 1  (MedChemExpress)
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Drp1 inhibits mitophagy by facilitating HK2 separation from the mitochondria. RMECs were pretreated with (A, B) INT-777 (30 μM) or <t>(C)</t> <t>Mdivi-1</t> (10 μM) for 2 h and then exposed with or without HG (33 mM) for 3 days. RMECs were pretreated with INT-777 (30 μM) or INT-777 (30 μM) + 3-BP (10 μM) for 2 h followed by HG (33 mM) for (D, E) three or (F) 5 days. (A, C) HK2 expression in mitochondrial and cytosolic fractions was detected by Western blot. All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. HG group. (B) DAPI, HK2 and TOMM20 were subjected to representative immunofluorescence analysis. S (s cale bar = 25 μm. (D) Western blot analysis showed the protein expression levels of PINK1 and Parkin. (E) JC-1 dye was used to assess mitochondrial membrane potential in different groups. Quantification of the ratio of fluorescence intensity (red/green) is shown in the bar chart. S (s cale bar = 25 μm ) . (F) Cell viability was measured using the CCK8 assay. All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. HG group, & $ p < 0.05 vs. INT-777 group.
Mdivi 1, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Drp1 inhibits mitophagy by facilitating HK2 separation from the mitochondria. RMECs were pretreated with (A, B) INT-777 (30 μM) or (C) Mdivi-1 (10 μM) for 2 h and then exposed with or without HG (33 mM) for 3 days. RMECs were pretreated with INT-777 (30 μM) or INT-777 (30 μM) + 3-BP (10 μM) for 2 h followed by HG (33 mM) for (D, E) three or (F) 5 days. (A, C) HK2 expression in mitochondrial and cytosolic fractions was detected by Western blot. All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. HG group. (B) DAPI, HK2 and TOMM20 were subjected to representative immunofluorescence analysis. S (s cale bar = 25 μm. (D) Western blot analysis showed the protein expression levels of PINK1 and Parkin. (E) JC-1 dye was used to assess mitochondrial membrane potential in different groups. Quantification of the ratio of fluorescence intensity (red/green) is shown in the bar chart. S (s cale bar = 25 μm ) . (F) Cell viability was measured using the CCK8 assay. All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. HG group, & $ p < 0.05 vs. INT-777 group.

Journal: Frontiers in Cell and Developmental Biology

Article Title: TGR5 Activation Ameliorates Mitochondrial Homeostasis via Regulating the PKCδ/Drp1-HK2 Signaling in Diabetic Retinopathy

doi: 10.3389/fcell.2021.759421

Figure Lengend Snippet: Drp1 inhibits mitophagy by facilitating HK2 separation from the mitochondria. RMECs were pretreated with (A, B) INT-777 (30 μM) or (C) Mdivi-1 (10 μM) for 2 h and then exposed with or without HG (33 mM) for 3 days. RMECs were pretreated with INT-777 (30 μM) or INT-777 (30 μM) + 3-BP (10 μM) for 2 h followed by HG (33 mM) for (D, E) three or (F) 5 days. (A, C) HK2 expression in mitochondrial and cytosolic fractions was detected by Western blot. All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. HG group. (B) DAPI, HK2 and TOMM20 were subjected to representative immunofluorescence analysis. S (s cale bar = 25 μm. (D) Western blot analysis showed the protein expression levels of PINK1 and Parkin. (E) JC-1 dye was used to assess mitochondrial membrane potential in different groups. Quantification of the ratio of fluorescence intensity (red/green) is shown in the bar chart. S (s cale bar = 25 μm ) . (F) Cell viability was measured using the CCK8 assay. All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. HG group, & $ p < 0.05 vs. INT-777 group.

Article Snippet: Mdivi-1, 3-bromopyruvic acid, INT-777, 3-methyladenine, Rapamycin, BAPTA-AM, and Z-VAD (OMe)-FMK were procured from MedChemExpress (NJ, United States).

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Membrane, Fluorescence, CCK-8 Assay

Mdivi-1 and Rapamycin alleviate the AAV8-shTGR5-induced retinal dysfunction in vivo . (A) Experimental workflow overview of in vivo experiments. (B) TGR5 shRNA lentivirus knockdown efficiency was assessed by Western blot. (C) Rats were infused with Evans Blue dye (3%) for 2 h. Red fluorescence dots indicate retinal vascular leakage in the flat-mounted retina by fluorescence microscopy. (D) Representative images of acellular capillaries were revealed by retinal trypsin digestion assay and quantified in random fields. ( S s cale bar = 25 μm ) . (E) Representative images for haematoxylin–eosin (HE) staining in rat retinas. Retinal thickness was evaluated in sections. ( S s cale bar = 25 μm ) . All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. AAV8-vector group. & $ p < 0.05 vs. AAV8-shTGR5 group.

Journal: Frontiers in Cell and Developmental Biology

Article Title: TGR5 Activation Ameliorates Mitochondrial Homeostasis via Regulating the PKCδ/Drp1-HK2 Signaling in Diabetic Retinopathy

doi: 10.3389/fcell.2021.759421

Figure Lengend Snippet: Mdivi-1 and Rapamycin alleviate the AAV8-shTGR5-induced retinal dysfunction in vivo . (A) Experimental workflow overview of in vivo experiments. (B) TGR5 shRNA lentivirus knockdown efficiency was assessed by Western blot. (C) Rats were infused with Evans Blue dye (3%) for 2 h. Red fluorescence dots indicate retinal vascular leakage in the flat-mounted retina by fluorescence microscopy. (D) Representative images of acellular capillaries were revealed by retinal trypsin digestion assay and quantified in random fields. ( S s cale bar = 25 μm ) . (E) Representative images for haematoxylin–eosin (HE) staining in rat retinas. Retinal thickness was evaluated in sections. ( S s cale bar = 25 μm ) . All data were presented as mean ± SD and analysed by RM ANOVA with Tukey’s HSD test. # p < 0.05 vs. control group, * p < 0.05 vs. AAV8-vector group. & $ p < 0.05 vs. AAV8-shTGR5 group.

Article Snippet: Mdivi-1, 3-bromopyruvic acid, INT-777, 3-methyladenine, Rapamycin, BAPTA-AM, and Z-VAD (OMe)-FMK were procured from MedChemExpress (NJ, United States).

Techniques: In Vivo, shRNA, Knockdown, Western Blot, Fluorescence, Microscopy, Trypsin Digestion Assay, Staining, Control, Plasmid Preparation