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Blood, Vol. 112, Issue 13, 5228-5237, December 15, 2008
Ribosomal protein S19 deficiency in zebrafish leads to developmental abnormalities and defective erythropoiesis through activation of p53 protein family
Blood Danilova et al.
112: 5228
Supplemental materials for: Danilova et al
Files in this Data Supplement:
- Table S1. Primers and PCR conditions (PDF, 23.6 KB)
- Figure S1. In contrast to erythropoiesis, development of myeloid and lymphoid lineages is less affected in RPS19-deficient zebrafish (JPG, 40.3 KB)
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(A, B) Myeloid cells expressing myeloperoxidase are present in RPS19-deficient zebrafish in the same numbers as in wild-type fish. 24 hpf, in situ hybridization with myeloperoxidase (Bennett 2001) probe. (C) Lymphoid cells are present in morphants’ thymus although in severely deformed embryos thymus architecture can be distorted and staining appears patchy. Day 5, in situ hybridization with recombination activating gene 1 (rag1) (Willett 1997) probe.
Bennett CM, Kanki JP, Rhodes J, et al. Myelopoiesis in the zebrafish, Danio rerio. Blood. 2001;98:643-651.
Willett CE, Cherry JJ, Steiner LA. Characterization and expression of the recombination activating genes (rag1 and rag2) of zebrafish. Immunogenetics. 1997;45:394-404.
- Figure S2. Expression of p53 family genes in embryos with mutations in ribosomal proteins S8, S11, and S18 (JPG, 48.1 KB)
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RNA was prepared from pool of 20–40 mutant embryos. Real-time PCR was performed using iQ SYBR Green Super Mix and a MyiQ Single-Color Real-Time PCR thermal cycler (Biorad, Hercules, CA). Levels of mRNA expression relative to wild-type embryos were normalized to β-actin mRNA and calculated according to the ΔCτ method.
- Figure S3. p53−/− zebrafish embryos injected with RPS19 morpholinos had no obvious morphological defects (JPG, 61.3 KB)
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(A) At 24 hpf, p53-deficient RPS19 morphants are very similar to control embryos, only slightly smaller. In contrast to wild-type morphants, they do not have prominent apoptosis in the brain (B). p53-deficient RPS19 morphants grow slower than controls and are smaller at 48 hpf. Their pigmentation is also delayed. (C) At day 5, p53-deficient morphants do not grow, stay small and some develop edema (arrow).
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