Correlation between TFAM-bound mtDNA amounts at the D-loop region and at the OriL region.

<p>A strong positive correlation was found between the TFAM-bound amounts of mtDNA at the assayed sub-regions in the joined animals from the AL-MA and CRO groups, but not in the AL-28 rats. The relative amounts of mtDNA immunoprecipitated by TFAM were determined by quantitative Real Time PCR (qRT-PCR) using primers for the D-loop and OriL sub-regions. Each sample was analyzed in triplicate. The calculation of the relative amount of TFAM-bound mtDNA was performed according to the formula 2^ΔCTx -2^ΔCTb, where ΔCTx is the C_T difference between the C_T values of the input and of the immunoprecipitated sample and ΔCTb is the C_T difference between the C_T values of the input and of the –Ab sample. Pearson’s tests were performed to determine whether the amounts of TFAM-bound mtDNA at both origins of replication correlate with each other. The correlation was highly significant (p<0.005, correlation coefficient: 0.952) in the AL-MA and CRO joined groups (n=10, filled line); the correlation was not statistically significant and reached a lower value (p= N.S., correlation coefficient: 0.409) in the AL-28 group (n=6, dashed line).