Supplementary Figure 8: Colony count and flow cytometric analysis of GM colonies.

(a) Comprehensive analysis of myeloid cell colonies. Data are from five experiments with eight litters: wild type n = 10, heterozygous n = 13, knockout n = 12. Numbers of colonies have been normalized to the input of 500 LSK cells. Center value, mean; error bars, s.e.m. P values were calculated by two-tailed unpaired t test. The F test when performed to compare the variances of the wild-type, heterozygous, and knockout groups showed, that variances differed significantly in total colonies (wild type vs. heterozygous, ***P < 0.001; wild type vs. knockout, ***P < 0.001), GM colonies (wild type vs. heterozygous, ***P < 0.001; wild type vs. knockout, **P = 0.003), M colonies (wild type vs. knockout, *P = 0.011), and G colonies (wild type vs. heterozygous, *P = 0.036; wild type vs. knockout, ***P < 0.001). Unpaired t tests with Welch’s correction showed that differences in means are significant in total colonies (wild type vs. heterozygous, *P = 0.037; wild type vs. knockout, **P = 0.004), in GM colonies (wild type vs. heterozygous, *P = 0.019; wild type vs. knockout, *P = 0.013), in M colonies (wild type vs. knockout, *P = 0.022), and in G colonies (wild type vs. knockout, *P = 0.011). This is likely to be due to sampling of experiments, outliers, and low numbers of colonies in M/G knockout samples. (b) Representative FACS plots of myeloid GM colonies derived from a showing expression of myeloid surface markers. Numbers are representative of the percentage of CD11b⁺Gr1⁺ and CD11b⁺Ly6c⁺ cells. CD11b⁺Gr1⁺ and CD11b⁺Ly6c⁺ cells are absent in Smarcd2^−/− colonies. Data shown are from one experiment (wild type n = 1, heterozygous n = 2, knockout n = 2), subsequently confirmed by a second experiment with wild type n = 5, heterozygous n = 4, and knockout n = 6, for which data are not shown. Replicates: 2.