mDia2 regulates stress-induced hematopoietic cell re-population.

(A) The composition of indicated colony forming units (CFU) determined based on morphology from primary plate seeded with 1×10⁴ BMMC cells from control or mDia2^fl/fl Vav-Cre mice. Data are presented as mean ± SEM, n = 2 in each group. (B) In vitro colony assays with serial plating. Representative phase contract views illustrating colony morphology under primary (1°), secondary (2°), and tertiary (3°) plating. (C) The CFU numbers in B were scored under a microscope. (D) The size of colonies from B was quantified by measuring the area of each unit with Image J. 1°: 162 colonies in control; 130 colonies in mDia2^fl/fl Vav-Cre; 2°:109 colonies in control; 99 colonies in mDia2^fl/fl Vav-Cre; 3°: 85 colonies in control; 40 colonies in mDia2^fl/fl Vav-Cre. Data were pooled from 3 mice in each group for C and D. (E) Kaplan-Meier survival curve depicting the survival of indicated mice challenged three times with 150 mg/kg 5-FU each time. The arrows indicate the experimental timelines for CBC and FACS assays. (F) Peripheral blood cell counts of mice in E determined on day 7. (G) Representative flow cytometric plots showing HSPC populations on day 21 after 5-FU treatment. (H) Cell cycle profiling of HSPCs from G. Error bars represent the SEM of the mean. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001, n.s., not significant. Two-tailed unpaired student’s t-test was used to generate the p values.