|
Fig. 2 foxn1/Casper Mutant Engrafts HSCs without Irradiation (A) Schematic representation of the workflow of HSC transplantation. CD41-GFPlo cells were sorted from the KM of Tg (CD41:GFP), and 1 × 104 CD41-GFPlo cells were injected into each recipient (Casper, 25-Gy-irradiated Casper, foxn1/Casper, and 15-Gy-irradiated foxn1/Casper). (B) The survival rate of each recipient at 60 days post transplantation (dpt). The irradiated foxn1/Casper showed a low survival rate after transplantation (data presented as days-percentage survival, t test; ∗p < 0.05, ∗∗p < 0.01; n (CD41-GFPlo > Casper) = 10, n (CD41-GFPlo > irradiated Casper) = 19, n (CD41-GFPlo > foxn1/Casper) = 23, and n (CD41-GFPlo > irradiated foxn1/Casper) = 18). (C) Imaging of representative recipients at 60 dpt. CD41-GFP+ cells could be visualized under a GFP field in the KM of nonirradiated foxn1/Casper mutant. The red dashed lines indicate the KM regions. n = 5; scale bar, 1 mm (kidney). (D) Confocal imaging of engrafted CD41-GFP+ cells at 60 dpt in tail region. White arrows indicate the CD41-GFPhi and CD41-GFPlo cells (top), and two magnified pictures show the CD41-GFPhi and CD41-GFPlo cells, respectively (bottom). Scale bar, 100 μm (top), 10 μm (bottom). (E) Representative flow-cytometric analysis of recipient KM at 60 dpt. The dots in the rectangles indicate engrafted CD41-GFP+ cells (mean ± SD, n [CD41-GFPlo > Casper] = 10, n [CD41-GFPlo > irradiated Casper] = 8, n [CD41-GFPlo > foxn1/Casper] = 8, and n [CD41-GFPlo > irradiated foxn1/Casper] = 4). (F) The statistical analysis of (E) (mean ± SD, one-way ANOVA, Tukey's; N.S., nonsignificant; ∗p < 0.05, ∗∗p < 0.01). See also Figure S2.