Characterization of novel miRNAs. (a) Sequence alignment of the novel miRNA candidates. Highly conserved 'seed' regions are highlighted in black and gray. Novel miRNAs are colored in red. (b) The expression of some novel miRNAs during development. Blue-colored and red-colored bars represent the results of quantitative RT-PCR and Solexa sequencing, respectively. The vertical axis indicates the relative expression level. The data were standardized to the expression in young adult hermaphrodites as 1. 'Ad' (young adult hermaphrodites) marked with an asterisk were cultured at 23°C, under the same condition as males, in order to rule out the possibility that male-enriched expression of these novel miRNAs is due to a higher culture temperature. Since Solexa sequencing was not performed for young adult hermaphrodites cultured at 23°C, this was shown as N.D. Error bars represent standard error. E, embryo; L, larval stage.