Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2015 Oct 2;290(46):27644–27659. doi: 10.1074/jbc.M115.654129

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

PMC Copyright notice

FIGURE 9. — NMNAT3 is present in mitochondria of human cells, and its expression level is unaffected by changes in the mitochondrial NAD⁺ content. A, Western blot detection of endogenous and overexpressed NMNAT3 in 293 cells with a monoclonal NMNAT3-specific antibody. The monoclonal antibody detected both the endogenous protein and the overexpressed FLAG-tagged recombinant NMNAT3. No cross-reactivity of the monoclonal antibody with NMNAT1 was observed. β-Tubulin served as a loading control. B, whole cell lysate (WCL), cytosolic and mitochondrial fractions from 293 cells were subjected to immunoblot analysis using the monoclonal NMNAT3-specific antibody. Superoxide dismutase (SOD2) served as a control for the purity of mitochondrial fraction, whereas β-tubulin served as a control for the cytosolic fraction. C, detection of endogenous NMNAT3 in lysates from 293 cell lines with altered mitochondrial NAD⁺ availability (293mitoPARP and 293AtNDT2), as well as control cells (293mitoEGFP and 293SLC25A32 and parental 293 cells). Lysates were subjected to immunoblot analysis using the NMNAT3-specific antibody. β-Tubulin served as a loading control. hNMNAT3, bacterially expressed, purified His₆-tagged human NMNAT3 (47).