Shortened microloader tip

Adult wild-type male zebrafish, close in age and weight (0.7 ± 0.05 g), were randomly chosen for the experiment and maintained as above. Prior to treatment, fish were anesthetized by gradual decrease in water temperature to 12°C. 10 μL of 10 mM EdU in 1X PBS was orally administrated to the anesthetized fish by gavaging the shortened microloader tip (Eppendorf) into the mouth and esophagus. EdU solution was delivered to the anterior portion of the foregut every 12 h and until the fish were euthanized (0.5–60 h after the first EdU pulse).

Vivo-Morpholinos (MOs) (Gene Tools, Philomath, OR) incorporate a knock-down sequence modified with a dendrimer delivery moiety. The prmt1 MOs’ sequence (5′-TGTCTGCCGTCTCCGCCATTTCGAT-3′) were published in previous studies (Tsai et al., 2011 (link)). The morpholino (MO) experiment was performed following the protocol published by Matsuda and Shi (2010) (link). Briefly, 2 nmol (4 μL of 0.5 mM stock) of translation blocking prmt1 Vivo-MO in PBS (PBS) was orally administrated to the anesthetized adult zebrafish by gavaging the shortened microloader tip (Eppendorf) into the mouth and esophagus. The prmt1 Vivo-MO was administered once per day for 4 days and the intestinal tissue was harvested 2 days later (day 6).