Nature research paper: Compartmentalized metabolism supports midgestation mammalian development
Whole embryos were collected from GD10.5 pregant mice into 1× PBS and mechanically disrupted using disposable pestles and then filtered through a 40-µM cell strainer to remove clumps. Antibody staining was performed for 20 min on ice, followed by washing with HBSS and centrifugation at 200for 5 min. Cells were stained with directly conjugated antibodies against mouse CD71 , mouse Ter119 , mouse CD41 and mouse CD117 .
Mice were allocated to experiments randomly and samples were processed in an arbitrary order, but formal randomization techniques were not used. Samples sizes were not pre-determined based on statistical power calculations but were based on our experience with these assays. For most experiments, the minimum number of mice was 3, with some exceptions where the embryo/placenta numbers were≥ 10.
Prior to analysing the statistical significance of differences among groups, we tested whether data were normally distributed and whether variance was similar among groups. To test for normality, we performed the Shapiro–Wilk tests when 3 ≤≥ 20. To test whether variability significantly differed among groups we performed-tests or Levene’s median tests .