Plasmid Preparation:Article Title: Enhanced Levels of Peroxisome-Derived H 2 O 2 Do Not Induce Pexophagy but Impair Autophagic Flux in HEK-293 and HeLa Cells
Article Snippet: .. The plasmid encoding po-mKeima (pMF2005) was generated by amplifying the corresponding cDNA via PCR (template: mKeima-Red-N1 (Addgene plasmid 54597; gift from Dr. M. Davidson (Florida State University, Tallahassee, FL, USA)); primers: 5′-ggg agatct accatggtgagcgtgatcgccaagcag-3’ and 5′-tcccc gcggccgc ttacagcttgctctt gcccagcagggagtggcgggcgatg-3’ (the restriction enzyme recognition sites and nucleotides encoding the prototypic C-terminal PTS1 “-KSKL” are underlined and indicated in bold, respectively)) and ligating the Bgl II/Not I-digested PCR product into Bgl II/Not I-restricted pEGFP-N1 (Clontech, Mountain View, CA, USA; 6085-1). .. The plasmid encoding OPTN-GFP was obtained from Addgene (plasmid 27052; gift from Dr. B. Yue (University of Illinois, Chicago, IL, USA) [ ]).
Article Title: Enhanced Levels of Peroxisome-Derived H 2 O 2 Do Not Induce Pexophagy but Impair Autophagic Flux in HEK-293 and HeLa Cells.
Article Snippet: .. The plasmid encoding po-mKeima (pMF2005) was generated by amplifying the corresponding cDNA via PCR (template: mKeima-Red-N1 (Addgene plasmid 54597; gift from Dr. M. Davidson (Florida State University, Tallahassee, FL, USA)); primers: 5′- gggagatctaccatggtgagcgtgatcgccaagcag-3’ and 5′-tccccgcggccgcttacagcttgctcttgcccagcagggagtggcgggcgatg-3’ (the restriction enzyme recognition sites and nucleotides encoding the prototypic C-terminal PTS1 “-KSKL” are underlined and indicated in bold, respectively)) and ligating the Bgl II/Not I-digested PCR product into Bgl II/Not I-restricted pEGFP-N1 (Clontech, Mountain View, CA, USA; 6085-1). .. The plasmid encoding OPTN-GFP was obtained from Addgene (plasmid 27052; gift from Dr. B. Yue (University of Illinois, Chicago, IL, USA) [36]).
Generated:Article Title: Enhanced Levels of Peroxisome-Derived H 2 O 2 Do Not Induce Pexophagy but Impair Autophagic Flux in HEK-293 and HeLa Cells
Article Snippet: .. The plasmid encoding po-mKeima (pMF2005) was generated by amplifying the corresponding cDNA via PCR (template: mKeima-Red-N1 (Addgene plasmid 54597; gift from Dr. M. Davidson (Florida State University, Tallahassee, FL, USA)); primers: 5′-ggg agatct accatggtgagcgtgatcgccaagcag-3’ and 5′-tcccc gcggccgc ttacagcttgctctt gcccagcagggagtggcgggcgatg-3’ (the restriction enzyme recognition sites and nucleotides encoding the prototypic C-terminal PTS1 “-KSKL” are underlined and indicated in bold, respectively)) and ligating the Bgl II/Not I-digested PCR product into Bgl II/Not I-restricted pEGFP-N1 (Clontech, Mountain View, CA, USA; 6085-1). .. The plasmid encoding OPTN-GFP was obtained from Addgene (plasmid 27052; gift from Dr. B. Yue (University of Illinois, Chicago, IL, USA) [ ]).
Article Title: Enhanced Levels of Peroxisome-Derived H 2 O 2 Do Not Induce Pexophagy but Impair Autophagic Flux in HEK-293 and HeLa Cells.
Article Snippet: .. The plasmid encoding po-mKeima (pMF2005) was generated by amplifying the corresponding cDNA via PCR (template: mKeima-Red-N1 (Addgene plasmid 54597; gift from Dr. M. Davidson (Florida State University, Tallahassee, FL, USA)); primers: 5′- gggagatctaccatggtgagcgtgatcgccaagcag-3’ and 5′-tccccgcggccgcttacagcttgctcttgcccagcagggagtggcgggcgatg-3’ (the restriction enzyme recognition sites and nucleotides encoding the prototypic C-terminal PTS1 “-KSKL” are underlined and indicated in bold, respectively)) and ligating the Bgl II/Not I-digested PCR product into Bgl II/Not I-restricted pEGFP-N1 (Clontech, Mountain View, CA, USA; 6085-1). .. The plasmid encoding OPTN-GFP was obtained from Addgene (plasmid 27052; gift from Dr. B. Yue (University of Illinois, Chicago, IL, USA) [36]).
Polymerase Chain Reaction:Article Title: Enhanced Levels of Peroxisome-Derived H 2 O 2 Do Not Induce Pexophagy but Impair Autophagic Flux in HEK-293 and HeLa Cells
Article Snippet: .. The plasmid encoding po-mKeima (pMF2005) was generated by amplifying the corresponding cDNA via PCR (template: mKeima-Red-N1 (Addgene plasmid 54597; gift from Dr. M. Davidson (Florida State University, Tallahassee, FL, USA)); primers: 5′-ggg agatct accatggtgagcgtgatcgccaagcag-3’ and 5′-tcccc gcggccgc ttacagcttgctctt gcccagcagggagtggcgggcgatg-3’ (the restriction enzyme recognition sites and nucleotides encoding the prototypic C-terminal PTS1 “-KSKL” are underlined and indicated in bold, respectively)) and ligating the Bgl II/Not I-digested PCR product into Bgl II/Not I-restricted pEGFP-N1 (Clontech, Mountain View, CA, USA; 6085-1). .. The plasmid encoding OPTN-GFP was obtained from Addgene (plasmid 27052; gift from Dr. B. Yue (University of Illinois, Chicago, IL, USA) [ ]).
Article Title: Enhanced Levels of Peroxisome-Derived H 2 O 2 Do Not Induce Pexophagy but Impair Autophagic Flux in HEK-293 and HeLa Cells.
Article Snippet: .. The plasmid encoding po-mKeima (pMF2005) was generated by amplifying the corresponding cDNA via PCR (template: mKeima-Red-N1 (Addgene plasmid 54597; gift from Dr. M. Davidson (Florida State University, Tallahassee, FL, USA)); primers: 5′- gggagatctaccatggtgagcgtgatcgccaagcag-3’ and 5′-tccccgcggccgcttacagcttgctcttgcccagcagggagtggcgggcgatg-3’ (the restriction enzyme recognition sites and nucleotides encoding the prototypic C-terminal PTS1 “-KSKL” are underlined and indicated in bold, respectively)) and ligating the Bgl II/Not I-digested PCR product into Bgl II/Not I-restricted pEGFP-N1 (Clontech, Mountain View, CA, USA; 6085-1). .. The plasmid encoding OPTN-GFP was obtained from Addgene (plasmid 27052; gift from Dr. B. Yue (University of Illinois, Chicago, IL, USA) [36]).
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