Recombinant mouse amphiregulin

Pancreatic islets were isolated from NOD.scid mice as described above. Whole islets were then cultured in RPMI supplemented with 10% Fetal Bovine Serum (FBS) for 7 h. Additional stimulation conditions included recombinant mouse Areg (100 ng/mL) (Recombinant mouse Amphiregulin, Carrier-free, Biolegend), Thapsigargin (5 uM) (Enzo Life Sciences), or mAreg plus Thapsigargin. After 7 h, total RNA was extracted following the manufacturer’s protocol from Qiagen RNeasy kit. First strand cDNA synthesis was performed using the manufacturer’s protocol from High-Capacity Reverse Transcriptase kit (Applied Biosciences). The following targets were analyzed by qPCR (Insulin, Grp78, sXBP1, CHOP, ATF4, ATF6, GAPDH) using a QuantStudio6 (Applied Biosystems). Primers for each target are listed in Table 1.Table 1

Primer sequences of targets used in qPCR.

Target		Sequence
Insulin	Forward	GTCAAGCAGCACCTTTGTGGTTCC
	Reverse	ACAATGCCACGCTTCTGCTG
Grp78	Forward	TGCTGCTAGGCCTGCTCCGA
	Reverse	CGACCACCGTGCCCACATCC
sXbp1	Forward	GAGTCCGCAGCAGGTGC
	Reverse	CAAAAGGATATCAGACTCAGAATCTGAA
Atf4	Forward	GCCGGTTTAAGTTGTGTGCT
	Reverse	CTGGATTCGAGGAATGTGCT
Atf6	Forward	GATGCAGCACATGAGGCTTA
	Reverse	CAGGAACGTGCTGAGTTGAA
Chop	Forward	CGGAACCTGAGGAGAGAGTG
	Reverse	CGTTTCCTGGGGATGAGATA
Gapdh	Forward	TGCACCACCAACTGCTTAG
	Reverse	GGATGCAGGGATGATGTTC

Pancreatic islets were isolated from NOD.scid mice as described above. Whole islets were then cultured in RPMI supplemented with 10% Fetal Bovine Serum (FBS) for 7 hrs. Additional stimulation conditions included recombinant mouse Areg (100ng/mL) (Recombinant mouse Amphiregulin, Carrier-free, Biolegend), Thapsigargin (5uM) (Enzo Life Sciences), or mAreg plus Thapsigargin. After 7 hrs, total RNA was extracted following the manufacturer’s protocol from Qiagen RNeasy kit. First strand cDNA synthesis was performed using the manufacturer’s protocol from High-Capacity Reverse Transcriptase kit (Applied Biosciences). The following targets were analyzed by qPCR (Insulin, Grp78, sXBP1, CHOP, ATF4, ATF6, GAPDH) using a QuantStudio6 (Applied Biosystems). Primers for each target are listed in Table 1.