Asymmetric LysoBrite inheritance predicts CD49c, CD34, and CD71 production. (A) Experimental design. (B) Dividing CD49fHSC with simultaneous quantification of LysoBrite and CD49c dynamics. Representative video frames of HSCs with a-/symmetric LysoBrite inheritance (trees 1 and 3 in panel C, respectively). Cells with symmetric LysoBrite inheritance produce comparable levels of CD49c in both daughters. With asymmetric inheritance, Lyso^Low daughters receive less CD49c and produce more CD49c throughout their cell cycle (green arrowhead, also in panel C). n = 5 independent experiments. Scale bar: 10 µm. (C) Representative quantification of LysoBrite inheritance and CD49c expression dynamics in paired daughter cells. Lyso^Low daughters produce more CD49c. Fold sister differences early after division are indicated, respectively. (D) Heatmap and clustering (top) and cluster frequency (bottom) of paired daughter cell CD49c dynamics after the a-/symmetric inheritance of LysoBrite. Each row represents a CD49fHSC daughter pair (#1 and #2). Cluster 1: asymmetric (mean paired daughter difference 1472 ± 159); Cluster 2: symmetric (91 ± 112). Lyso^Low (daughter #2) produces more CD49c. Lower left, mean CD49c fluorescence intensities of a-/symmetric paired daughter cell clusters, mean ± SD. n = 5 independent experiments. (E) CD33, CD201, CD71, TMRM, CD49c, or CD34 production after a-/symmetric LysoBrite inheritance in paired daughter cells. Asymmetric Lyso^Low daughters (white) produce more CD49c, CD34, and CD71 than their sisters (black). Comparable CD33, CD201, and TMRM production in daughter cells after asymmetric LysoBrite inheritance. Two-tailed Mann-Whitney test. n = 4 independent experiments for CD71, TMRM, and CD34, n = 5 for CD33, CD201, and CD49c.