Mitochondrial Targeting of Lig1 Protein

Rabbit anti-FUS (Cat# A300-302A) and anti-Lig1 (A301-136A) antibodies were procured from Bethyl Laboratories, Inc. Mouse anti-FLAG antibody (A8592) was obtained from Sigma-Aldrich, and mouse anti-Lig3 antibody (Cat# ab587) was purchased from Abcam. Mouse anti-Tom20 (SC-17764), anti-HSP60 (SC-13115) and anti-PCNA (SC-56) antibodies were procured from Santa Cruz Biotechnology. Fluorescent secondary antibodies, Alexa Fluor 488 anti-mouse (Cat# A28175), and Texas Red anti-rabbit antibody (Cat# T-2767) were obtained from Life Technologies. The antibodies were diluted at 1:1000 for western blotting, 1:500 for immunofluorescence and 1:100 for PLA.
The human Lig1 coding sequence was re-cloned into pCDNA3.1 plasmid as an N-terminal FLAG and COX8 gene mitochondrial targeting sequence containing construct (FLAG-MTS-Lig1) ^{53 (link)}.

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Kodavati M., Wang H., Guo W., Mitra J., Hegde P.M., Provasek V., Maloji Rao V.H., Vedula I., Zhang A., Mitra S., Tomkinson A.E., Hamilton D.J., Bosch L.V, & Hegde M.L. (2023). FUS Unveiled in Mitochondrial DNA Repair and Targeted Ligase-1 Expression Rescues Repair-Defects in FUS-Linked Neurodegeneration. Research Square.

Publication Preprint 2023

Rabbit anti-FUS anti-Lig1 (A301-136A) Mouse anti-FLAG antibody (A8592) mouse anti-Lig3 antibody anti-PCNA (SC-56) Alexa Fluor 488 anti-mouse Texas Red anti-rabbit antibody

A301 Alexa fluor 488 Anti antibody Antibodies Coding sequence Human Immunofluorescence Lig1 Lig3 Mitochondrial Mouse Pcna Plasmid Rabbit

Corresponding Organization : Cornell University

Other organizations : Allen Institute for Brain Science, KU Leuven, Texas A&M University, BioEnergetics (United States), Methodist Hospital, New Mexico Cancer Center, University of New Mexico, VIB-KU Leuven Center for Brain & Disease Research

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Variable analysis

independent variables

Expression of FLAG-MTS-Lig1 construct

dependent variables

Protein levels of FUS, Lig1, Lig3, Tom20, HSP60, and PCNA

control variables

Antibody dilutions for western blotting (1:1000), immunofluorescence (1:500), and PLA (1:100)

controls

Positive control: None specified
Negative control: None specified

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