TRIzol reagent was employed to extract miR-27b-3p and PPARG from the H9C2 cell line. The TaKaRa PrimeScript 1st Strand cDNA Synthesis Kit (TaKaRa Bio Inc, Japan) was adopted to reverse-transcribe miR-27b-3p and PPARG into cDNA. SYBR Green qPCR Master Mix (MedChemExpress, NJ, USA) and the IQ5 Multicolor qRT-PCR Detection System (Bio-Rad, USA) were utilized for real-time fluorescent quantitative PCR. The relative expression was calculated based on the 2−ΔΔCT value of each gene, and all experiments were repeated 3 times. GAPDH and U6 were taken as the internal parameters of PPARG and miR-27b-3p, respectively [23 (link)]. The primer sequences are detailed in Table 1 .
The sequence of each molecular primer
| Genes | Primer sequences (5’‐3’) | |
|---|---|---|
| PPARG | Forward | TTTCCTGTCAAGATCGCCCT |
| Reverse | TTGCAGTGGGGATGTCTCAT | |
| miR-27b-3p | Forward | AGGGTTCACAGTGGCTAAG |
| Reverse | GAGAGGAGAGGAAGAGGGAA | |
| GAPDH | Forward | TGTGTCCGTCGTGGATCTGA |
| Reverse | TTGCTGTTGAAGTCGCAGGAG | |
| U6 | Forward | CTCGCTTCGGCAGCACA |
| Reverse | AACGCTCTCACGAATTTGCGT | |
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