Journal: Nucleic Acids Research
Article Title: Stable nuclear expression of ATP8 and ATP6 genes rescues a mtDNA Complex V null mutant
doi: 10.1093/nar/gkw756
Figure Lengend Snippet: OxPhos complexes reassemble upon exogenous expression of ATP8 and ATP6 proteins in the A8/A6 mut cell line. Mitochondria-enriched fractions (∼25 μg protein/lane) from WT (lane 1), A8/A6 mut + A8F + A6F (lane 2), A8/A6 mut + A8F (lane 3) and A8/A6 mut (lane 4) were electrophoresed on 4–12% Bis-Tris BN-PAGE gels. Proteins were transferred onto PVDF membranes and immunodetected with the following antibodies: ( A ) panel i: anti-ATP8, ii: anti-FLAG, iii: anti-ATP5h, iv: anti-ATP5O (OSCP), and ( B ) panel i: anti-NDUFS4 (Complex I), ii: anti-SDHB (Complex II), iii: anti-Core 2 protein (Complex III), iv: anti-MT-CO2 (Complex IV). Protein standards are indicated on the left. * Immunodetected protein band, ** CV dimers, + degraded products of Complex III.
Article Snippet: The following antibodies were used in this study: Anti-FLAG (Cat# F1804) from Sigma, St. Louis, MO; anti-UQCRC2 : Complex III Core2 protein (Cat# MS304) and ATP5h (Cat# MS504) from MitoSciences, Eugene, OR; NDUFS4 (Cat# AP6932b); NDUFS3 from GeneTex, Irvine, CA (Cat# GTX113504); V5 from Genscript (Cat# A01724), and SDHB (Cat# AP19974b) from Abgent, San Diego, CA; ATP6 from mAbdx (Clone# 1G7-1G2); MT-CO2 (Cat# A303-600A) from Bethyl Laboratories, Montgomery, TX and ATP5O/OSCP (Cat# TA804572) from Origene Technologies, Rockville, MD, USA.
Techniques: Expressing