Expression of gene transcripts encoding COPII components is reduced in feelgood. (A) Schematic of zebrafish Creb3l2 expression constructs. ‘X’ in the basic domain marks the feelgood N301K mutation. (B) Luciferase reporter assays. Human fibroblasts were transfected with the indicated expression constructs and the firefly luciferase reporter containing 0.8 kb of the zebrafish sec23a promoter. Renilla luciferase control plasmid was also used as transfection efficiency control. Luciferase activities were compared with those of cells transfected with empty vector to determine fold induction. Red line indicates baseline activity adjusted to Renilla luciferase values. (C,D) qPCR analysis of the fold expression change of COPII-related transcripts creb3l2, sec23a, sec23b, sec24d and sec24c (C), and ER-stress-related transcripts s1p, bip and sil1 (D) in feelgood, crusher (sec23a) and bulldog (sec24d) mutants compared with wild types. RNA was extracted form whole embryos at 80 hpf. All results are normalized to β-actin and then to wild type. *P<0.05; **P<0.005.
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