TABLE 2.

Real-time PCR conditions utilized for mtDNA and cDNA quantification^a

Gene	Sense primer	Antisense primer	MgCl2	Temp
MT-RNR1	TAGCCCTAAACCTCAACAGT	TGCGCTTACTTTGTAGCCTTCAT	5	62
RNR1	ATCCTTCGATGTCGGC	AGCACATACACCAAATGTCT	6	60
ACTB	CGACATGGAGAAAATCTGGC	CCATCCTGCGTCTGGACCT	2.5	60
DDIT3	GCCAAAATCAGAGCTGGAAC	TCTTGCAGGTCCTCATACCA	3.5	60
MT-CO2	TACGGCGGACTAATCTTCAA	AAAACAGATGCAATTCCCGG	2.5	60
COX4I1	TGGATGAGAAAGTCGAGTTG	TCGTTATCATGTGGCAGAAG	2.5	60
CYBB	AATGCCCAATCCCTCAGTTT	TCCCACTTCCATTTTGAACC	3.5	58
NOX4	TCCGTTGGTTTGCAGATTTAC	CAAAAGTTTCCACCGAGGAC	3	58
NRF1	GCAACAGGAAAGAAACGGAA	ATTGTCCTCTGTATCTCACC	2.5	60
GABPA	TGCTGCACTGGAAGGCTATA	ACCGATATAGACCTCACCACA	2.5	60
TFAM	TGTGTATTTACCGAGGTGGT	TTAGAAGAATTGCCCAGCGT	2.5	60
PPARGC1B	CTGAAGATGACGTGGGT	TGATGTTGGGTAGGATGTG	3.5	60
PPARGC1A	AAGAGCGCCGTGTGATTTAT	ACCTACCGTTATACCTGTGA	2.5	60
PPRC1	CCACGTACTCAGGGTT	GGTCTAGGGGCCTCTT	3	60
UCP2	CAGGATCGGCCTCTACGAC	GTCAACGGTTCGGTTGTTGC	3	60

^aFor each amplification, serial dilutions of linearized plasmid vector pGEM-T Easy (Promega) containing the target gene as its insert were used as standards. All standard plasmids were purified with Nucleobond AX (Macherey-Nagel), linearized, and quantified by fluorometry using Sybr green I and λ phage DNA cut with HindIII as the standard (Invitrogen). Magnesium concentration is expressed as mmol/liter, and annealing temperature as °C.