Transfection of Hippocampal Neurons and Neuro-2a Cells

Hippocampal neurons were transfected at 7 DIV using Lipofectamine 2000 (Thermo Fisher, Cat# 11668019). Experiments were carried out after 20–24 hr. The pEGFP-C1 plasmid is from Clontech (Addgene plasmid # 13031, RRID:Addgene_13031), the plasmids for EGFP-Rab10 WT (RRID:Addgene_49472), EGFP-Rab10 T23N (RRID:Addgene_49545), and EGFP-Rab10 Q68L (RRID:Addgene_49544) were a gift from Marci Scidmore (Huang et al., 2010 (link)), TrkB-FLAG plasmid was a gift from Francis Lee (Chen et al., 2005 (link)). Neuro-2a cells were transfected using Lipofectamine 3000 (Thermo Fisher, Cat# L3000001) and the experiments were done 48 hr later. The GFP-KIF13B plasmid (RRID:Addgene_134626) was a gift from Marvin Bentley (Yang et al., 2019 (link)), whilst the HA-Rab10 plasmid was provided by Dario Alessi and Miratul Muqit (Dundee University, DU44250).

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Lazo O.M, & Schiavo G. (2023). Rab10 regulates the sorting of internalised TrkB for retrograde axonal transport. eLife, 12, e81532.

Publication 2023

Lipofectamine 2000 pEGFP-C1 plasmid Lipofectamine 3000

Cells Lipofectamine Lipofectamine 2000 Neurons Plasmid Trkb

Corresponding Organization :

Other organizations : National Hospital for Neurology and Neurosurgery, UK Dementia Research Institute, University College London

Top 5 similar protocols

Variable analysis

independent variables

Transfection of hippocampal neurons with pEGFP-C1, EGFP-Rab10 WT, EGFP-Rab10 T23N, EGFP-Rab10 Q68L, and TrkB-FLAG plasmids
Transfection of Neuro-2a cells with GFP-KIF13B and HA-Rab10 plasmids

dependent variables

Not explicitly mentioned

control variables

Hippocampal neurons transfected at 7 DIV
Experiments carried out after 20-24 hr of transfection
Neuro-2a cells transfected and experiments done 48 hr later

controls

Positive control: pEGFP-C1 plasmid
Negative controls: Not explicitly mentioned

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