Fig. 3. Flow cytometric analysis of whole embryo cell suspensions. (A,B) Flow cytometric quantification of total cell number (unbroken line) and 95% confidence interval (broken lines) from pooled wild-type (A, n=65) and Igf2^+m/-p (B, n=84) embryos dated by seminal plugs and using a quadratic line of best fit (Minitab). Inserts show linear plots of log_e cell number versus age (days). (C) Box plots of normalised (100%) cell number (subtracting sub-G1) of each embryo relative to the mean cell number of the respective genotype in each litter (box=interquartile range, vertical line=95% confidence interval, horizontal line=median). Insert shows pooled values. (D) Ratios of mean cell numbers per litter for Igf2^+m/-p (-p) over wild-type (+p) embryos matched per litter and aged by the mean cell number of wild-type embryos in each litter (blue diamonds, n=15 litters, 140 embryos, P=0.005 at E11, Student’s t-test). Line shows linear regression of all data points. (E) Somite number plotted against log_e cell number for wild type (blue, n=65) and Igf2^+m/-p (red, n=84). (F) Ratios of mean somite numbers and line of best fit for Igf2^+m/-p (-p) over wild type (+p) embryos matched per litter and aged by the mean cell number of wild-type embryos in each litter (n=10 litters, 124 embryos). Litters with at least two embryos of each genotype were used.

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