Fig. 1

Bioinformatic and Expression Analysis of Zebrafish rpl22 and rpl22l1 (A–D) Sequence alignment of RNA-binding domains, phylogenetic analysis, and synteny analysis. (A) Red-boxed area denotes RNA-binding motif. (B) Numbers on the branches of the phylogenetic tree are bootstrap values. (C) The human (1p36) and zebrafish rpl22 (LG11) loci are flanked by a common set of genes (CHD5, PLEKGH5, PHF13, VAMP3, PER3, PARK7, ERRF11, and SLC45A1). (D) Human (3q26) and zebrafish rpl22l1 (lg24) loci are flanked by a common set of genes (MYNN, LRRC34, LRRC31, SKIL, CLDN11, EIF-5A2, and TNFSF10). Orthologous RPL22 and RPL22L1 gene symbols are in red and bold. (E–F2) Zebrafish rpl22 and rpl22l1 WISH analysis on 24 hpf embryos. (E and F) Lateral views with head to the left. (E2 and F2) Insets depict the ICM region delimited by dashed lines. (G) Real-time PCR quantification of rpl22 and rpl22l1 mRNA levels in hematopoietic progenitor populations. Tg fish lines were employed to identify the following progenitors: (1) heart, cmcl2:EGFP at 2 dpf, (2) T cell, lck:EGFP at 5 dpf, (3) myeloid, mpo:EGFP at 5 dpf, (4) erythroid, gata1:dsRED2 at 5 dpf, and (5) HSC, cd41:EFGP at 3.5 dpf. GFP+ cells were isolated by flow cytometry, after which rpl22 and rpl22l1 expression was measured by real-time PCR. Results of triplicate measurements were normalized to b-actin and whole embryo homogenates and represented graphically as the mean ± SD. The results were combined from at least three separate experiments with representative photographs depicting the observed phenotypes. Unless otherwise specified, all images represent lateral views with the head to the left. See also Figure S1.