Total RNA samples were isolated from the whole organoids using the RNeasy Mini Kit. 1 μg of RNA was used to generated cDNA using the iScript Select cDNA Synthesis Kit. Real time quantitative PCR was performed using the SsoFast EvaGreen Supermix in the CFX96 Real-Time PCR System. The PCR cycling conditions were: 95°C f or 15 min, followed by 40 two-step cycles at 94°C for 10 sec and 60°C for 45 sec. Primers used w ere as follows: NKX2-1 forward: 5′-GAGTCCAGAGCCATGTCAGC-3′, reverse:5′- GCATAAAACAGCTTTGGGGTGT-3′; SST forward: 5′-GCTGCTGTCTGAACCCAAC-3′, reverse: 5′-CGTTCTCGGGGTGCCATAG-3′; PV forward: 5′-AAAGAGTGCGGATGATGTGAAG-3′, reverse:5′-ACCCCAATTTTGCCGTCCC-3′; NPY forward: 5′-CGCTGCGACACTACATCAAC-3′, reverse: 5′-CAGGGTCTTCAAGCCGAGTT-3′; CALB forward: 5′-TCCAAGCAGTAGACATGCTGTT-3′, reverse: 5′-ACAACCATAGAACTGTCCCACA-3′.
