Fig. 1

A: Flowchart depicting cold and hot temperature treatments in zebrafish for sampling. Three-month-old zebrafish reared at 28 °C were subjected to cold treatment by exposure to 18 °C for 12 h and 8 °C for 12 h. Hot temperature treatment was carried out by exposing fish to 38 °C for 4 h, with controls maintained at 28 °C. Samples for qRT-PCR and immunoblot analysis were collected at three temperature-time points (28 °C, 38 °C 4 h, 8 °C 12 h). Sample size: n=30. B: Dusp1 expression under cold and hot treatment measured using qRT-PCR compared with normal temperature controls. Data are mean±SD. ^**: P<0.01; ^***: P<0.001. C: Western blot analysis of DUSP1 protein levels in gills under thermal stress, with ACTB as the loading control. D: Schematic of targeted dusp1 gene editing by CRISPR/Cas9 and disruption of dusp1 at exon 1. E: Western blot validation of loss of dusp1 in dusp1-KO fish indicated by two tissues examined.