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. 2012 Mar 24;40(13):6097–6108. doi: 10.1093/nar/gks257

Figure 5.

Figure 5.

Recombinant C4orf14 binds to DNA and to RNA non-specifically. (A–D) Gel-retardation assays using 53 nM radiolabelled 170-bp DNA incubated with different amounts of recombinant C4orf14 as indicated, without (A), or pre-mixed with 2.8 -µM TFAM (B) or 5.6 µM TFAM (C) for 20 min at 37°C. The boxed lanes each contained the same amount of C4orf14 (14 µM). (D) Radiolabelled mtRNA was incubated without protein or with 3.5, 7, 14 or 28 µM C4orf14-MBP for 20 min at 37°C, prior to separation by 1D-AGE. (E) C4orf14-MBP captured mtRNA (see ‘Materials and methods’ section) was end-labelled and incubated with a Southern blot of amplified fragments of mtDNA corresponding to 12S and 16S rRNAs, cytochrome c oxidase subunit II (COX2) and NADH dehydrogenase subunit 1 (ND1). Ribosomal RNAs are of much greater abundance than mRNAs, which accounts for the disparity in signal between the two types of gene.