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1) Product Images from "The MFN1 and MFN2 mitofusins promote clustering between mitochondria and peroxisomes"
Article Title: The MFN1 and MFN2 mitofusins promote clustering between mitochondria and peroxisomes
Journal: Communications Biology
Figure Legend Snippet: Exogenous expression of MFNs enhance the PerMit Venus reporter signal. a Schematic for the constructs of Cyto-V(N), Mito-V(N), and Po-V(C). Cyto-V(N), HA tag fused to the N terminus of Venus with a linker composed of 4 × GGSG (indicated with blue box); Mito-V(N), Tom20 fused to the N terminus Venus with a HA tag and two linkers as indicated; Po-V(C), Myc tag and PEX26(residues 237-305) fused to the C terminus of Venus (residues 155-238). b Immuno-blots for Cyto-V(N), Mito-V(N), and Po-V(C). c Immunofluorescence images of Cyto-V(N), Mito-V(N), and Po-V(C) co-stained with mitochondrial COX4 and peroxisomal ABCD3 in HeLa cells. Scale bars, 5 μm. d Immunofluorescence images of Venus co-stained with mitochondrial COX4 and peroxisomal ABCD3 in PerMit Venus and control cells. Scale bars, 5 μm. e Immunofluorescence images of PerMit Venus cells with exogenously expressed MFNs. PerMit Venus cells were transfected with empty vector, MFN1-FLAG, or MFN2-FLAG plasmids for 36 h. FLAG (Alexa Fluor 568) and peroxisomal membrane protein PEX14 (Alexa Fluor 647) were immunostained. Scale bars, 5 μm. f Integrated density of ( e ), Vector, n = 67; MFN1-FLAG, n = 73; MFN2-FLAG, n = 72. *** p
Techniques Used: Expressing, Construct, Western Blot, Immunofluorescence, Staining, Transfection, Plasmid Preparation
Figure Legend Snippet: Exogenous expression of MFN induces peroxisome/mitochondrion clustering. Immunofluorescence images of overexpressed MFN-EGFP. HeLa cells were transfected with free EGFP ( a ) or EGFP fused MFN ( b ) plasmids for 36 h. Peroxisomal matrix protein catalase (Alexa Fluor 555) and peroxisomal membrane protein ABCD3 (Alexa Fluor 647) were immune-stained. Scale bars, 5 μm. c Peroxisomal membrane protein PEX14 (Alexa Fluor 555) and outer mitochondrial membrane protein Tom20 (Alexa Fluor 647) were immunostained with or without exogenously expressed MFNs. Scale bars, 5 μm. d Immuno-blots of overexpressed MFN1-EGFP and MFN2-EGFP. 1.5 µg plasmids were transfected into HeLa cells in one well in a six-well cell culture plate for 36 h and immunoblotted with indicated antibodies. e Immunofluorescence images of overexpressed MFN2-EGFP and other organelle markers. HeLa cells were transfected with MFN2-EGFP and immunostained for peroxisomal membrane protein ABCD3 (Alexa Fluor 647) and other organelle markers (Alexa Fluor 555): calnexin (endoplasmic reticulum), EEA1 (early endosome), GM130 (Golgi), and LAMP1 (lysosome). Scale bars, 5 μm. f Immunofluorescence images of overexpressed MFN1-EGFP and other organelle markers stained the same as in ( c ).
Techniques Used: Expressing, Immunofluorescence, Transfection, Staining, Western Blot, Cell Culture