Figure 9. Asymmetric inheritance of p-p38^MAPK and numb in 2T WT-LSK-SLAM.

(a) LSK-SLAM cells from 0T, 2T WT and 2T-p190-B^−/− mice were cultured for 40 h and stained for p-p38^MAPK (in green), Numb (in red) and DAPI (inblue), scale bar, 10 μm. 3D plots using ImageJ software represent distributions of p-p38^MAPK and Numb in daughter cells. (b) Bar graphs represent per cent of symmetric and asymmetric distribution of p-p38^MAPK and Numb in mitotic cells. 35–40 mitotic cells from three independent experiments were analysed. P values were calculated by Fisher exact 2 × 2 contingency table comparing 2T-WT to control and 2T-p190-B^−/− to 2T-WT. (c) Effect of p38^MAPK inhibitor on daughter cell multilineage differentiation potential. Single 2T-WT LSK-SLAM cells were cultured with SCF+TPO for one division. Paired-daughters were separated and individually cultured with serum and multiple cytokines for mature differentiation for 14 days in the presence or absence of p38^MAPK inhibitor. Resultant clones were analysed as in Fig. 1 (n=20 pairs in two independent experiments). P values were calculated by Fisher exact 2 × 2 contingency table by comparing per cent of symmetric and asymmetric divisions of p38^MAPK inhibitor versus DMSO.