Fig. 3

(A) Northern analysis of RNA isolated from WT and tti^s450 larvae at 120 hpf using 5′ETS, ITS1, and ITS2 probes to detect precursor forms of rRNA. Elf1α is a loading control. a-d correspond to the rRNA intermediates depicted in Figure 3B. (B) Schematic diagram showing the rRNA processing pathway in zebrafish [60]. The sites of hybridization of the 5′ETS, ITS1 and ITS2 probes are indicated. (C) Representative E-Bioanalyser analysis of total RNA isolated from WT and tti^s450 larvae at 120 hpf demonstrates a reduction in the 18S peak in tti^s450 larvae resulting in an elevated 28S/18S rRNA ratio in tti^s450 (D). Graphical representation of the experiment shown in C. Data are represented as mean +/ SD (n = 5). (E) Representative polysome fractionation analysis performed on WT and tti^s450 larvae at 96 hpf demonstrates reduced levels of 40S ribosomal subunits and 80S monosomes and an increase in free 60S subunits in tti^s450 larvae compared to WT. (F) Graphical representation of the experiment shown in E. Data are represented as mean +/ SD (n = 5) *p<0.05.