Table 1.

qRT-PCR primers used during late stages of β-cell differentiation

Gene	Sequence (5’ to 3’)	Function
GAPDH	Forward: AAGGTGAAGGTCGGAGTCAA Reverse: AATGAAGGGGTCATTGATGG	Housekeeping gene used to normalize gene expression
PDX1	Forward: CCTTTCCCATGGATGAAGTC Reverse: CGTCCGCTTGTTCTCCTC	Pancreatic and duodenal homeobox 1; expression begins during posterior foregut stage and remains high5,6,36
NKX6.1	Forward: GGGGATGACAGAGAGTCAGG Reverse: CGAGTCCTGCTTCTTCTTGG	Homeobox 1 protein NK6; expression begins during pancreatic progenitor stage and indicates differentiation into monohormonal cells6,37
NEUROD1	Forward: GCCCCAGGGTTATGAGACTAT Reverse: ATCAGCCCACTCTCGCTGTA	Basic helix-loop-helix transcription factor; endocrine precursor marker involved in β-cell differentiation and expansion33
NGN3	Forward: CTATTCTTTTGCGCCGGTAG Reverse: CTTCGTCTTCCGAGGCTCT	Neurogenin 3, member of basic helix-loop-helix transcription factor family; proendocrine gene critical for determination of endocrine cell specification36
PCSK1	Forward: CTCTGGCTGCTGGCATCT Reverse: CGGGTCATACTCAGAGGTCC	Gene encoding proprotein convertase 1 involved in pro-insulin processing29
MafA	Forward: GAGAGCGAGAAGTGCCAACT Reverse: TTCTCCTTGTACAGGTCCCG	V-maf musculoaponeurotic fibrosarcoma oncogene homolog A; maturation marker expressed late in β-cell maturation36,38
ECAD	Forward: TTGACGCCGAGAGCTACAC Reverse: GACCGGTGCAATCTTCAAA	E-cadherin; cell surface adhesion protein involved in intercellular communication between β-cells25
Insulin	Forward: TTCTACACACCCAAGACCCG Reverse: CAATGCCACGCTTCTGC	β-cell hormone
Glucagon	Forward: TGCTCTCTCTTCACCTGCTCT Reverse: AGCTGCCTTGTACCAGCATT	α-cell hormone