Rab5-Mediated Vesicle Dynamics Protocol

pET-GST-R5BD-hRABEP1 was purchased from Vector Builder. Rab5a-pmCherryC1 was a gift from Christien Merrifield (Addgene plasmid #27679; http://n2t.net/addgene:27679, accessed on 31 May 2019; RRID:Addgene_27679) [41 (link)]. pcDNA4-Vps34-Flag was from Qing Zhong (Addgene plasmid #24398; http://n2t.net/addgene:24398, accessed on 12 November 2018; RRID:Addgene_24398) [42 (link)]. mCherry-Rab5CA(Q79L) was from Sergio Grinstein (Addgene plasmid #35138; http://n2t.net/addgene:35138, accessed on 26 April 2018; RRID:Addgene_35138) [43 (link)]. TagRFP-T-EEA1 was from Silvia Corvera (Addgene plasmid #42635; http://n2t.net/addgene:42635, accessed on 9 November 2017; RRID:Addgene_42635) [44 (link)]. pTag-BFP-C-h-Rab5a-c-Myc was from James Johnson (Addgene plasmid #79801; http://n2t.net/addgene:79801, accessed on 9 November 2017; RRID:Addgene_79801) [45 (link)]. RFP-Rab5 140 and 180 mutants were generated with primers (5′-GCAGCAGCTGTTGACTTCCAGG-3′ and 5′-ATTTGCTAAGTCAGCTTTGTTTCCTGAC-3′) and (5′-GCAGCGCTGCCAAAGAATGAAC-3′ and 5′-AGCTATTGCCATAAATATTTCATTTACATTCATTG-3′), respectively, using a KOD-plus mutagenesis kit (Toyobo, Osaka, Japan).

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Mashimo M., Morozumi A., Nobeyama A., Kanzaki M., Negi S., Kato J., Moss J., Nomura A, & Fujii T. (2022). Poly(ADP-ribose) Polymerase 1 Mediates Rab5 Inactivation after DNA Damage. International Journal of Molecular Sciences, 23(14), 7827.

Publication 2022

pcDNA4-Vps34-Flag mCherry-Rab5CA(Q79L) pTag-BFP-C-h-Rab5a-c-Myc KOD-plus mutagenesis kit

Mutagenesis Plasmid Primers Vector

Corresponding Organization : Doshisha Women's College of Liberal Arts

Other organizations : National Heart Lung and Blood Institute, National Institutes of Health

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Variable analysis

independent variables

PET-GST-R5BD-hRABEP1 plasmid from Vector Builder
Rab5a-pmCherryC1 plasmid from Christien Merrifield
PcDNA4-Vps34-Flag plasmid from Qing Zhong
MCherry-Rab5CA(Q79L) plasmid from Sergio Grinstein
TagRFP-T-EEA1 plasmid from Silvia Corvera
PTag-BFP-C-h-Rab5a-c-Myc plasmid from James Johnson
RFP-Rab5 140 and 180 mutants generated using primers and a KOD-plus mutagenesis kit

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: Not mentioned
Negative controls: Not mentioned

Annotations

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