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. Author manuscript; available in PMC: 2013 May 4.
Published in final edited form as: Cell Stem Cell. 2012 May 4;10(5):520–530. doi: 10.1016/j.stem.2012.04.007

Figure 2. Increased Cdc42 activity correlates with a de-polarized phenotype in LT-HSCs.

Figure 2

(A), Representative distribution of Cdc42, tubulin and pericentrin-2 (Per-2) in young LT-HSCs determined by IF. Pictures are shown on a dark background (panels i, iii, v and vii) or as overlap with the phase contrast picture (panels ii, iv, vi and viii). Bar = 5µm. Panel ix and x show Cdc42 (red) and Per-2 (blue) distribution over the phase contrast picture. Panel xi: schematic presentation of a representative distribution of Cdc42 in young LT-HSCs (Per-2, blue dot; Cdc42, red dots). The arrow indicates the direction from “a” to “b” followed for determining fluorescence intensity in panel xii. Panel xii: representative fluorescence intensity plot obtained by collecting pixel intensity through the section of the cell as indicated in xi. (B), Representative distribution of Cdc42, tubulin and Per-2 in aged LT-HSCs determined by IF. Pictures are shown on a dark background (panels i, iii, v and vii) or as overlap with the phase contrast picture (panels ii, iv, vi and viii). Bar = 5µm. Panel ix shows Cdc42 (red) and Per-2 (blue) distribution over the phase contrast picture. Panel x: schematic presentation of a representative distribution of Cdc42 in aged LT-HSCs (Per-2, blue dot; Cdc42, red dots). The arrow indicates the direction from “a” to “b” followed for determining fluorescence intensity in panel xi. Panel xii: representative fluorescence intensity plot obtained by collecting pixel intensity through the section of the cell as indicated in x. (C), Percentage of young and aged LT-HSC cells with a polar distribution of Cdc42 and tubulin. Shown are mean +1 S.E., n=10; ~500–700 cells scored per sample in total. * p < 0.001. (D), Percentage of Per-2 polarized cells of Cdc42 polarized young and aged LT-HSCs. Cdc42-polarized cells were analyzed for Per-2 localization and scored positive when Per-2 was found at the Cdc42-pole. Shown are mean +1 S.E., n=4, ~150–250 cells scored per sample in total. * p < 0.05. (E), Representative distribution of Cdc42 and tubulin in young Cdc42GAP+/+ and Cdc42GAP−/− LT-HSCs. Pictures are shown on a dark background (panels i–vi) or as overlap with the phase contrast picture (panels vii and viii). Bar = 5µm. (F), Percentages of young Cdc42GAP+/+ (WT Control) and Cdc42GAP−/− LT-HSC cells with a polar distribution of Cdc42 and tubulin. Shown are mean mean +1 S.E., n=4, ~200–300 cells scored per sample in total. ** P < 0.01, *P < 0.05. See also Figure S2 and Movies S1–S3.