Fig. 3. Profile of mRNA expression for the V-ATPase B-subunit, as observed by northern blot analysis of total RNA isolated via (A) CsCl cushion or (B) RNeasy affinity spin column. Data for the 3500 bp message are plotted as circles with a broken line. Data for the 2250 bp message from post-diapause embryos are plotted as diamonds and a solid line, while that for diapause embryos are plotted as a cross. Two of the 4 h samples for plot B were excluded from analysis, as they formed insoluble precipitates during the drying step preceding suspension in gel loading buffer. For northern blots, values are expressed as means ± S.E.M. (N=3). Shared lowercase letters above error bars indicate that no significant difference exists between time points, as determined by Tukey's test. An asterisk denotes that a significant difference was found between two time points, as determined by the Nemenyi test. (C) Relative abundance of four A. franciscana developmental stages during incubation at 22–23°C. For each time point, a total of 332–631 individuals was counted, and the data were plotted as a percentage of the total.