MCAD. Medium chain acyl-coenzyme A dehydrogenase. AGGGTTTAGTTTTGAGTTGACGG. CCCCGCTTTTGTCATATTCCG. MCD. Malonyl-CoA decarboxylase.
SUPPLEMENTARY DATA
Supplementary Table 1. qRT-PCR Primers for Quantification of mRNA Gene Expression.
Gene COX-1 Cytochrome c oxidase subunit I COX-2 Cytochrome c oxidase subunit I CS Citrate synthase CytC Cytochrome C ERRa Estrogen receptor related receptor α MCAD Medium chain acyl-coenzyme A dehydrogenase MCD Malonyl-CoA decarboxylase MHCI Myosin heavy chain 1 mtTFA Mitochondrial transcription factor A
POLG-2 Mitochondrial DNA polymerase subunit gamma-2 SDH Succinate dehydrogenase Sirt-1 Sirtuin 1 SSBP-1 Stranded DNA binding protein 1 Tropn-1 Troponin I UCP-1 Uncoupling protein 1 UCP-2 Uncoupling protein 2
GAGGCGCTGGTAGACCTCT GAGGCTAGGTTCTTTCGTAACTC GAA CTG CAC ACA GAC CTG C GAC TGG GTT AAG CCA ATG CTC GCTGACGACTTCGACGACG TCGGTCAACAGGAGGTTGTCT CAACAAATGAGATGTGGCGATCA ACGAGCTTCTTACCAGCTATGA CCAGCACCTTCAGCTTCAGGTCCTTGAT TGCCGGAAGTTGAGAGGAAATCCAAGA T AGGCTTCCAGTACCATTAGGT CTGAGTGAGGCAAAGCTGATTT ATGGTTGGTTTCAAGGCCACA CGGTATCCAGAGGGAAAGTGAT
Supplementary Table 2. Primers for mtDNA Determination.
Primers 5’-3’ CCGCAAGGGAAAGATGAAAGAC-3’ mtDNA specific 5’-TCGTTTGGTTTCGGGGTTTC-3’ nuclear specific GCCAGCCTCTCCTGATTTTAGTGT GGGAACACAAAAGACCTCTTCTGG Gene
Supplementary Table 3. miRNA Probe Sequences for RNase protection assays.
Supplementary Figure 1. Transfection of miR-712-3p mimic did not alter mitochondrial biogenesis in cultured myotubes. C2C12 myotubes were transfected with NS or miR-712-3p mimic. Total RNA was isolated after 24 h transfection. A: NAD+ levels in myotubes were determined by calorimetric method. B mRNA expression of genes involved in mitochondrial biogenesis in the total RNA was examined by RT-qPCR. Each bar indicates mean ± SEM (n=3). White and black bars indicate NS (non-specific) and miR-712-3p mimic, respectively.