Apela is upregulated during zebrafish caudal fin regeneration. (A) Schematic representation of experimental plan showing amputation site and regenerated fin areas. Adult zebrafish were anesthetized in tricaine and fins were amputated and allowed to regenerate for indicated time periods. (B) Total RNA was isolated from fins (15–20 fins per time point) with uncut fins as controls (0 dpa) and analyzed by real-time PCR using specific primers for Apela or β-actin. Results are shown in the bar graph and are expressed as the ratio of the indicated transcripts relative to control (0 dpa). Results are shown as means ± S.E. of three experiments performed in triplicate. (C) Representative confocal images of control fins (0 dpa) and at 3 dpa (n = 12) subjected to immunofluorescence of Apela (red signal). (D) Quantification of fluorescence intensity in ImageJ. (E) Total RNA was isolated from fins (15–20 fins per time point) with uncut fins as controls (0 dpa) and analyzed by real-time PCR as in (A), using specific primers for Apln. (F) Aplnra and Aplnrb expression analysis by RT-PCR in uncut fins. (G) Aplnra expression analysis as in (A). The mean ± S.E values are shown. *p < 0.05. Scale bars represent 0.5 mm.
|
