Fig. 5: SRF restores engraftment defects in mDia2-deficient HSPCs.
a Quantitative analysis of the flow cytometry detected surface expression levels of beta2 integrins in c-Kit+ HSPCs from mDia2fl/fl or mDia2fl/flVav-Cre (cKO) mice transduced with indicated genes. N= 3 mice in each group, except mDia2 ΔDAD and mDia2 GBD/DID groups. b Restoration of the mRNA expression of beta2 integrins and indicated SRF downstream genes by overexpression of SRF in c-Kit+ cells from mDia2fl/flVav-Cre (cKO) mice. The experiments were performed in triplicate. N = 3 mice in each group. c Viral transduced c-kit+ HSPC from mDia2fl/fl or mDia2fl/flVav-Cre (cKO) mice were applied for in vitro adhesion assay with fibronectin precoated coverslip for 1 h at 37 °C. Adherent cells were randomly scored from N = 7 fields in mDia2fl/fl+Vector, N = 9 fields in cKO+Vector, N = 8 fields in cKO+SRFΔ5, and N = 6 fields in cKO+SRF-FL. Results are expressed as the adhesion ability relative to control cells transduced with vector. d Schematic diagram showing bone marrow transplantation experiments in e–g. e Chimerism studies of peripheral blood mononuclear cells derived from c-Kit+ cells that were transduced with the indicated constructs after competitive transplantation with CD45.1+ cells at indicated timepoints. f Quantification of multi-lineage reconstitutions in the peripheral blood from e at 3-month. g LSK and long-term HSC engraftment analyzed in bone marrow from 6-month-old transplanted mice shown in e. h Kaplan–Meier survival curve of indicated groups of mice after secondary transplantation illustrated in d. Error bars represent the SEM of the mean. p values compared to cKO+Vec group from g–j are presented on the right table. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Two-tailed unpaired Student’s t test was used to generate the p values. Significance for survival analyses was calculated by log-rank (Mantel–Cox) test.
