Live imaging of endogenous speck formation in the Tg(asc:asc-EGFP) line. Time-lapse imaging of a keratinocyte with a speck. Single plane merged with the brightfield is shown. (A) Yellow arrowheads highlight a second cell that appears to surround the speck-containing cell. Full time-lapse is included in Video 1. Live imaging of specks in the dorsal epidermis (B) and ventral fin (C) of 3-dpf Tg(asc:asc-EGFP) larvae. Merge with brightfield plane shows each speck is within a cell with altered morphology (dashed yellow line). (D) Tg(asc:asc-EGFP) 3-dpf larvae were treated with 25 µM CuSO4 for 1 h. At 1 and 3 h after treatment, the number of specks per larva were quantified (one-way ANOVA; ****, P < 0.0001). Live imaging of untreated and treated larvae showing high damage of epidermis and increase in specks (E), examples in treated embryo of single cells displaying altered morphology with and without speck formation (E′). Bars: (full larvae) 300 µm; (all others) 30 µm.
