ZFIN ID: ZDB-FIG-150929-14
Lippok et al., 2014 - Pou5f1 protein expression and posttranslational modification during early zebrafish development. Developmental dynamics : an official publication of the American Association of Anatomists   243(3):468-77 Full text @ Dev. Dyn.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Anatomical Term:
Stage Range: Sphere to Prim-5

Fig. 5

Temporal Pou5f1 profile during early embryo development. A: Pou5f1 protein amounts (green, around 60 kDa) from 4 hr postfertilization (hpf) to 24 hpf wild-type (WT) whole embryo extracts analyzed by fluorescent Western blot analysis. α-tubulin is shown as loading control (red, 55 kDa). In the Western blot, an additional faint higher molecular weight band is detected at approximately 150 kDa from mid-gastrula on, and a second band appears from the end of gastrulation at approximately 100 kDa. These bands are also present in MZspg embryos, and thus do not correspond to Pou5f1 forms but are nonspecific bands. B: Relative quantification of Pou5f1 immunoreactive band intensity from 4 to 24 hpf indicates that Pou5f1 protein amount is highest between 6 hpf (65% epiboly stage) and 10 hpf (100% epiboly stage). Pou5f1 band intensity values were normalized using the α-tubulin band intensity values. The Pou5f1 protein signal in WT at 90% epiboly stage was set to 100%. C: Relative quantification of pou5f1 mRNA message from 4 hpf (sphere) to 10 hpf (bud) by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). pou5f1 mRNA amount is highest at 50% epiboly stage and decays afterward. Elongation factor alpha (eef1a1a) was used as a normalization control. pou5f1 mRNA signal in WT at 50% epiboly stage was set to 100%. D: Pou5f1 at pre-MBT stages. Pou5f1 protein is already present in unfertilized eggs, but the band is shifted to higher molecular mass compared with a new Pou5f1 band that appears faintly at two-cell stage and then prominently from eight-cell stage on. The right part shows absence of Pou5f1 bands from MZspg mutant embryos. Fluorescent Western blot analysis using P2 Pou5f1 antibody (green) and anti-α-tubulin as loading control (red, 55 kDa).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
pou5f3 Ab2-pou5f3 WT standard conditions Sphere whole organism WB
Ab2-pou5f3 Dome whole organism WB
Sphere to Bud whole organism RTPCR
Ab2-pou5f3 Shield whole organism WB
Ab2-pou5f3 50%-epiboly whole organism WB
Ab2-pou5f3 90%-epiboly whole organism WB
Ab2-pou5f3 Bud whole organism WB
Ab2-pou5f3 1-4 somites whole organism WB
Ab2-pou5f3 Prim-5 Not Detected whole organism WB
Antibody Labeling Details No data available
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal Developmental dynamics : an official publication of the American Association of Anatomists for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Dev. Dyn.