Table 1.

Primers and annealing temperatures used for RT-PCR and bisulfite sequencing

Gene	Sequence (5′ → 3′)	Product size (bp)	Annealing temperature (°C)
RT-PCR
β-actin-F	ACACCTTCTACAATGAGCTG	816	56
β-actin-R	CTGCTTGCTGATCCACATCT
Runx2-F	CCTGAACTCTGCACCAAGTC	234	55
Runx2-R	GAGGTGGCAGTGTCATCATC
Col1a1-F	TGGACGCCATCAAGGTCTACTGC	455	56
Col1a1-R	GGAGGTCTTGGTGGTTTTGTATTCG
Osterix-F	CCTCTGCGGGACTCAACAAC	355	56
Osterix-R	TGCCTGGACCTGGTGAGATG
Osteocalcin-F	CAGACAAGTCCCACACAGCAGC	165	56
Osteocalcin-R	TGTTCACTACCTTATTGCCCTCC
Bisulfite sequencing
Dlx5 region1-F	TTAGTAGGTAGGAAATAATGGG	707	52
Dlx5 region1-R	CGATTCTTAATACTCTTTTCTTACT
Dlx5 region1-F	TGGAGTGGATGTAGGTAATG (nested)	446	55
Dlx5 region1-R	TATAAACTAAAAAACAATTAAACAC (nested)
Dlx5 region2-F	GAGTTATGATAGGAGTGTTTGAT	365	50
Dlx5 region2-R	CCTACCTAACTAACTAATAACACT
Dlx5 region2-F	TGTTTGATAGAAGAGTTTTAAGTATT (nested)	183	56
Dlx5 region2-R	AAACAAAAATAAAAAAACAATAACC (nested)