HNF-4α determines hepatic differentiation of human mesenchymal stem cells from bone marrow

Table 1 Primer sets used for reverse transcription-polymerase chain reaction analysis

Gene	Forward primer	Reverse primer	Product size (bp)
Alb	TGCTTGAATGTGCTGATGACAGGG	AAGGCAAGTCAGGGCATCTCATC	161
AFP	TGCAGCCAAAGTGAAGAGGGAAGA	CATAGCGAGCAGCCCAAAGAAGAA	216
TAT	TGAGCAGTCTGTCCACTGCCT	ATGTGAATGAGGAGGATCTGAG	358
G6P	GCTGGAGTCCTGTCAGGCATTGC	TAGAGCTGAGGCGGAATGGGAG	350
TO	ATACAGAGACTTCAGGGAGC	TGGTTGGGTTCATCTTCGGTATC	299
HNF-1α	GTGTCTACAACTGGTTTGCC	TGTAGACACTGTCACTAAGG	251
HNF-3β	CACCACTACGCCTTCAACC	GGTAGTAGGAGGTATCTGCGG	235
HNF-4	CTGCTCGGAGCCACAAAGAGATCCATG	ATCATCTGCCACGTGATGCTCTGCA	371
C/EBPα	CAAGAAGTCGGTGGACAAGAAC	CCTCATCTTAGACGCACCAAGT	450
β2M	TGACGTGTGAACCATG	TGGAGACAGCACTCAAAG	242

The thermo-cycling parameters for β2M: 94°C for 5 min/28 cycles of 94°C for 30 s, 56°C for 30 s, and 72°C for 30 s/72°C for 10 min; The thermo-cycling parameters for other genes: 94°C for 5 min/38 cycles of 94°C for 30 s, 56-58°C for 30 s, and 72°C for 30 s/72°C for 10 min. Alb: Albumin; AFP: α-fetoprotein; TAT: Tyrosine-aminotransferase; TO: Tryptophan 2,3-dioxygenase; G6P: Glucose 6-phosphatase; β2M: β-2-microglobulin..

Table 2 Primer sets for real-time reverse transcription-polymerase chain reaction analysis

Gene	Forward primer	Reverse primer	Probe
HNF-1α	TGAGTCCGGGCTTCACAC	GGCTGCTGGAGGACACTG	42
HNF-3β	GCACCTGCAGATTCTGATTTT	GACTTCCCTGCAACAACAGC	25
HNF-4α	ATTGACAACCTGTTGCAGGA	CGTTGGTTCCCATATGTTCC	77
HNF-4γ	CTTGGTGGAATGGGCTAAA	AGCTCTCAACAGTGCCACCT	8
HNF-6	CCTGGAGCAAACTCAAATCC	TTCTTTCCTTTTGCATGCTG	88
C/EBPα	CAACACTTGTATCTGGCCTCTG	CGAGCAAAACCAAAACAAAAC	3
PPARα	GTCCCCGCAGATTCTACATT	GAAGCTGTCCTGGCTCAGAT	14
Alb	TGTTGCCAAGCTGCTGATA	CCTTCATCCCGAAGTTCATC	27
TAT	TGCTGAGCAGTCTATCCACT	CTGCTCACAGAACTCCTGGAT	67
G6P	GCTGCTCATTTTCCTCATCAA	TTCTGTAACAGCAATGCCTGA	67
CYP1A1	GACAGATCCCATCTGCCCTA	CAAACTTGTGTCTCTTGTTGTGC	80
CYP1A2	CAAGAAATGCTGTGTCTTCGTAA	AGAGGGGTCCTCCCACAG	59
CYP2A6	CAAAAAGGACACCAAGTTTCG	AGAGCCCAGCATAGGGTACA	69
CYP2B6	GAAGCTTTTATCCCCTTCTCCT	GCCATGGAGAAGTTCTGGAG	35
CYP2C8	AAGAAAAGTGACTACTTCATGCCTTT	CAAGTCCTTCTCCTGCACAA	18
CYP2C9	ATTGACCTTCTCCCCACCA	CAGGGAAATTAATATGGTTGTGC	43
CYP2C19	GTCCAGAGATACATCGACCTCA	AGTGAGGGAAGTTAATATGGTTGTG	43
CYP2D6	AGGAGGAGTCGGGCTTTCT	CGCTGGGATATGCAGGAG	56
CYP2E1	CAAGCCATTTTCCACAGGA	CAACAAAAGAAACAACTCCATGC	67
HPRT	AGGTTGCAGCGAGGAAAAA	AGCCACCCTGGAAGAAACTT	59

The thermo-cycling parameters for real-time reverse transcription-polymerase chain reaction: 95°C for 10 min/50 cycles of 95°C for 10 s, 60°C for 30 s, and 72°C for 1 s/40°C for 30 s. Alb: Albumin; AFP: α-fetoprotein; TAT: Tyrosine-aminotransferase; G6P: Glucose 6-phosphatase; PPARα: Peroxisome proliferator-activated receptor α; HPRT: Hypoxanthine-guanine phosphoribosyltransferase.