Reproducibility of assays for quantifying polyadenylation of mitochondrial mRNA
| . | Control . | Anoxia . | Anoxia+1.5 mmol l−1 ATP . | Anoxia+low pH . | Low pH . |
|---|---|---|---|---|---|
| COXI | |||||
| Mean±s.e.m. | 8.23±0.24 | 1.09±0.06 | 0.84±0.02 | 1.02±0.03 | 1.16±0.07 |
| CV | 5.08 | 8.66 | 3.40 | 5.88 | 10.15 |
| ATP6 | |||||
| Mean±s.e.m. | 1.34±0.05 | 0.82±0.04 | 1.11±0.07 | 0.43±0.03 | 1.39±0.08 |
| CV | 5.59 | 7.21 | 9.80 | 12.30 | 9.54 |
| . | Control . | Anoxia . | Anoxia+1.5 mmol l−1 ATP . | Anoxia+low pH . | Low pH . |
|---|---|---|---|---|---|
| COXI | |||||
| Mean±s.e.m. | 8.23±0.24 | 1.09±0.06 | 0.84±0.02 | 1.02±0.03 | 1.16±0.07 |
| CV | 5.08 | 8.66 | 3.40 | 5.88 | 10.15 |
| ATP6 | |||||
| Mean±s.e.m. | 1.34±0.05 | 0.82±0.04 | 1.11±0.07 | 0.43±0.03 | 1.39±0.08 |
| CV | 5.59 | 7.21 | 9.80 | 12.30 | 9.54 |
ATP6, ATP synthase; COX1, cytochrome oxidase; CV, coefficient of variation.
N=3 PAT assays. At least two separate mitochondrial RNA preparations were used for each treatment. The values represent an average ratio of ethidium bromide fluorescence from the bottom (polyadenylated) bands of the PAT products to the top (non-adenylated) band, as measured with Molecular Dynamics Typhoon and ImageQuant software (see Materials and methods for details).