Fig. 4.

Deficiency of aPKCζ and aPKCλ does not affect interaction of HSC/P within the BM microenvironment. (A) Absolute numbers of CFU-Cs present in the peripheral blood of WT; Vav1-Cre or PKCζ^−/−; Vav1-Cre or aPKCλ^Δ/Δ; Vav1-Cre or aPKCζ^−/−λ^Δ/Δ; Vav1-Cre mice (n = 6–16 mice per group). Data represent average of two different experiments. (B) Experimental set up. 3 × 10⁶ BMNCs CD45.2⁺ WT; Vav1-Cre or PKCζ^−/−; Vav1-Cre or aPKCλ^Δ/Δ; Vav1-Cre or aPKCζ^−/−λ^Δ/Δ; Vav1-Cre mice, were noncompetitively transplanted into lethally irradiated CD45.1⁺ B6.SJL^{Ptprca Pep3b/BoyJ} WT mice. CD45.2⁺ chimera was monitored in the peripheral blood of the recipient mice. (C) Absolute numbers of CD45.2⁺CD11b⁺-myeloid cells present in the peripheral blood of CD45.1⁺ B6.SJL^{Ptprca Pep3b/BoyJ} mice after 8 wk of transplantation from B (n = 6–9 mice per group). (D) Absolute numbers of CD45.2⁺B220⁺ B-lymphoid cells present in the peripheral blood of CD45.1⁺ B6.SJL^{Ptprca Pep3b/BoyJ} mice after 8 wk of transplantation from B (n = 6–9 mice per group). (E) Absolute numbers of CD45.2⁺CD3⁺ T-lymphoid cells present in the peripheral blood of CD45.1⁺ B6.SJL^{Ptprca Pep3b/BoyJ} mice after 8 wk of transplantation from B (n = 6–9 mice per group). (C–E) Error bars represent SD. (F) Experimental set up. The 2 × 10⁶ BM cells from CD45.2⁺ WT; Vav1-Cre or aPKCλ^Δ/Δ; Vav1-Cre or aPKCζ^−/−λ^Δ/Δ; Vav1-Cre mice were mixed with 2 × 10⁶ CD45.1⁺ B6.SJL^{Ptprca Pep3b/BoyJ} WT BM cells and competitively (1:1 ratio) transplanted into lethally irradiated primary mice. (G) Evolution of CD45.2⁺ chimera in the peripheral blood of recipient mice from F (n = 7–10 mice per group). (H) Adhesion (performed in triplicate) of WT; Vav1-Cre, PKCζ^−/−; Vav1-Cre, aPKCλ^Δ/Δ; Vav1-Cre or aPKCζ^−/−λ^Δ/Δ; Vav1-Cre HSC/Ps to fibronectin (CH-296) in vitro. Error bars represent SEM. (I) Migration (performed in triplicate) of WT; Vav1-Cre, PKCζ^−/−; Vav1-Cre, aPKCλ^Δ/Δ; Vav1-Cre or aPKCζ^−/−λ^Δ/Δ; Vav1-Cre HSC/Ps toward CXCL12 in vitro. (G, I) Error bars represent SEM.