Blood, Vol. 113, Issue 9, 2038-2046, February 26, 2009
Essential role of spi-1–like (spi-1l) in zebrafish myeloid cell differentiation
Blood Bukrinsky et al. 113: 2038
Supplemental materials for: Bukrinsky et al
Files in this Data Supplement:
- Figure S1. Lateral view of 48 hpf embryos (JPG, 43.6 KB) -
Red and blue arrows indicate mRNA expression in the YK and the CHT regions respectively of wild-type embryos. Probes for genes and morpholino used are as indicated. (A) Expression of spi-1l in wild type embryo and (B) in silent heart embryos. Lack of difference in spi-1l expression pattern between wild type and silent heart embryo indicates de novo production of spi-1l cells in the CHT.
- Figure S2 (JPG, 151 KB) -
(A) RT-PCR analysis of spi-1l morphant and wild-type embryos. Embryo genotype and PCR product are as indicated. Spi-1l MO causes mis-splicing of the spi-1l transcript as demonstrated by a shorter transcript in spi-1l morphant embryo. (−) RT is a control reaction without the reverse transcriptase enzyme added to RT-PCR reaction. Beta actin is used as a loading control. (B–G) Image of the entire embryo shown in figure 3 (B–G). Lateral view of 28 hpf embryos. Red and blue arrows indicate mRNA expression in the YK and the CHT regions respectively of wild-type embryos. Probes for genes and morpholino used are as indicated. (A) Expression of mpx and (B) lcp1 in wild-type embryos. Spi-1l and pu.1 morphant embryos exhibit an almost complete absence of granulocyte (mpx) (C,E) and macrophage (lcp1) (D,F) markers respectively. (H) Expression of lcp1 and (I) mpx in wild-type embryos. Spi-1l AUG-MO morphant embryos exhibit an almost complete absence of lcp1 (J) and mpx (K) markers respectively. (L) Expression of mpx and (M) lcp1 in wild-type embryos. Control MO morphant embryos exhibit wild type expression of mpx (N) and (lcp1) (O) markers respectively.
- Figure S3 (JPG, 120 KB) -
(A–I) Lateral view of embryos. Red and blue arrows indicate mRNA expression in the A-LPM and the ICM regions respectively of wild-type embryos. Probes for genes and morpholino used are as indicated. (A) Wild-type expression of runx-1. (B) Expression of runx-1 is unchanged in spi-1l morphants. (C) Expression of runx-1 is slightly decreased in the ICM of pu.1 morphats. (D,F,H) Wild-type expression of c-myb (D), scl (F), and gata-2 (H). (E,G,I) Expression of c-myb (E), scl (G), and gata-2 (I) is unchanged in spi-1l morphants. (J,K) Ventral view of 5 dpf embryo. (J) Wild type expression of rag1 and (K) rag1 expression in spi-1l morphant embryo is unchanged.
- Figure S4 (JPG, 142 KB) -
(A–C) Image of the entire embryo shown in figure 4 (A–C). (A–C) Lateral view of the yolk (YK) region at 36 hpf of cloche wild-type sibling, cloche, and injected cloche embryo respectively. (A) Expression of spi-1l in a wild-type sibling from a cloche intercross. (B) Expression of spi-1l is absent in cloche mutant embryo. (C) Partial rescue of spi-1l expression in cloche mutant injected with pu.1 mRNA. (D–I) Image of the entire embryo shown in figure 4 (D–I). Etsrp as an upstream component of spi-1l and spi-1. Lateral view of embryos. Probes for genes, morpholino, and mRNA used are as indicated. Red and blue arrows indicate mRNA expression on the YK and in the CHT regions respectively of wild-type embryo. (D) Spi-1l and (E) pu.1 expression in control un-injected embryos at 28 hpf. Etsrp morphants lack spi-1l (F) and pu.1 (E) expression. Ectopic etsrp mRNA expression causes increase in the number of spi-1l (H), and pu.1 (I) cells.
- Figure S5 (JPG, 86 KB) -
(A–F) Image of the entire embryo shown in figure 5 (A–F). Lateral view of 26 hpf embryos. Probes for genes, morpholino, and injection mixture used are as indicated. Red and blue arrows indicate mRNA expression in the YK region and the CHT respectively. Wild-type expression of lcp1 (A) and mpx (B) in un-injected embryos. Spi-1 MO eliminates lcp1 (C) and mpx (D) expression. Embryos co-injected with pu.1 MO and spi-1l mRNA recover some lcp1 (E) and mpx (F) expression.
- Figure S6 (JPG, 103 KB) -
(A–F) Image of the entire embryo shown in figure 6 (A–F). Lateral view of the yolk (YK) region of embryos from a heterozygous cloche intercross at 36 hpf. Probes for genes, embryo genotype and mRNA injected are as indicated. Red arrows indicate mRNA expression in the YK region of a wild-type sibling from a cloche intercross. Wild-type sibling of cloche mutants express mpx (A) and lcp1 (B). Cloche mutant embryo lack expression of mpx (C) and lcp1 (D). Cloche embryos injected with spi-1l mRNA partially recover mpx (E) and lcp1 (F) expression.