Rat anti sox2

Embryos and culture cells were fixed with 4% paraformaldehyde (PFA) at room temperature (for 15 min for cells and up to E7.5 stage embryos, 60 min for E9.5 and E12.5). Whole E6.5 embryos were stained with rabbit anti-Eomes antibody (Abcam, Cat. No. ab23345). Blastocysts were stained with rabbit anti-GFP (Invitrogen, Cat. No. A-11122) and rat anti-Sox2 (eBioscience, Cat. No. 14-9811-82). After the fixation, E9.5 and E12.5 embryos were incubated with 20% sucrose/phosphate buffered saline (PBS) overnight at 4 °C, then embedded in OCT cryo-embedding compound (Sakura, Cat. No. 4583). Sections were stained with rat anti-Sox2 (eBioscience) and fluorescein isothiocyanate (FITC)-conjugated mouse anti-cTnT antibodies (Abcam, Cat. No. ab105439) (E9.5 only). Nuclei were stained with DAPI. Embryos and sections were imaged by either a Leica SP5 or Zeiss LSM880 confocal microscope. E7.5 PGC were stained with anti-Tfap2c (Santa Cruz, Cat. No. sc-8977), anti-Stella (Abcam, Cat. No. ab19878), and rat anti-Sox2 (eBioscience, Cat. No. 14-9811-82). FS cells were stained with rabbit anti-cMyc antibody (Abcam, Cat. No. ab30272).