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. 2021 Oct 28;31(8):1230–1241. doi: 10.1093/hmg/ddab314

Figure 3.

Figure 3

The longer tricistronic transcript of MT-ATP8, MT-ATP6 and MT-CO3 associates with the mitochondrial ribosome. (A) Immunoblotting of sucrose density gradient fractions of the cultured human fibroblasts with the indicated MT-ATP6 genotypes to profile the sedimentation profile of the mitochondrial ribosome. (B) Northern blotting of RNA isolated from the indicated sucrose density gradient fractions. (C) Left, a diagram of the poison primer extension assay to determine the tricistronic transcript from processed MT-CO3 in the indicated MT-ATP6 genotypes. Middle, a representative poison primer extension analysis from total RNA collected from the indicated sucrose gradient fractions. Right, quantification of the primer extension analysis from three independent experiments (mean ± SD) with the indicated MT-ATP6 genotypes. (D) Semi-quantitative RT-PCR of RNA isolated from the indicated sucrose gradient fractions of cultured human fibroblasts with the indicated MT-ATP6 genotypes. Left, a diagram of the assay. Right, representative image of the analysis. -, PCR with no cDNA template; -RT, PCR without the RT step. All data are representative of independent experiments.