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. 2013 Feb 1;24(3):184–193. doi: 10.1091/mbc.E12-09-0651

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© 2013 Fung et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 5: — Transient siRNA-mediated knockdown of MRPL14 in control fibroblasts phenocopies the C7orf30 knockdown. MRPL14 was knocked down using a Stealth RNAi construct, and a BLOCK-iT-Alexa Fluor oligonucleotide was used as a control. (A) SDS–PAGE immunoblot analysis of mitochondrial lysates using antibodies against MRPL14, C7orf30, OXPHOS proteins, and MRPs as indicated. (B) SDS–PAGE immunoblot analysis of isokinetic sucrose gradients loaded with mitochondrial lysates from Alexa control or siRNA-MRPL14 (si-L14) probed with antibodies against MRPS as indicated. (C) Pulse-chase labeling of newly synthesized mitochondrial polypeptides (at right; ND, subunits of complex I; COX, subunits of complex IV; cyt b, subunit of complex III; ATP, subunits of complex V) in control and siRNA-MRPL14 cells. (D) Quantification of the amount of labeled newly synthesized mitochondrial polypeptides in shRNA-C7orf30-A2-C2 and siRNA-MRPL14 fibroblast lines expressed as a percentage of their respective controls.