Total RNA from GMSCs was extracted by using TRIzol (Invitrogen). The levels of miR-4651 in GMSCs were detected by using a Hairpin-it microRNA and U6 snRNA Normalization RT-PCR Quantitation Kit (GenePharma, Suzhou, China). Two-microgram aliquots of RNA were reverse-transcribed into cDNA with random hexamers or oligo(dT) and reverse transcriptase according to the manufacturer’s instructions (Invitrogen). Then, real-time PCR was performed as previously described.54 (link) GAPDH and U6 were used to normalize mRNA and miRNA levels, respectively. Primer sequences are listed in Table 1 .
Primers sequences used in the real-time RT-PCR
| Gene symbol | Primer sequence (5′-3′) |
|---|---|
| GAPDH-F | CGGACCAATACGACCAAATCCG |
| GAPDH-R | AGCCACATCGCTCAGACACC |
| HMGA2-F | ACCCAGGGGAAGACCCAAA |
| HMGA2-R | CCTCTTGGCCGTTTTTCTCCA |
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