Figure 6. Reduced NF-κB Signaling and IKKβ Deficiency Leads to Loss of HSPC and Impaired Hematopoietic Reconstitution.

(A) Expression of NF-κB target genes in VavCre⁺ and Traf6^−/−VavCre⁺ LSK cells as determined by RNA-seq.

(B) Expression of select NF-κB target genes measured by qRT-PCR in Traf6^−/−VavCre⁺ relative to VavCre⁺ LSK cells.

(C) Immunoblot analysis of phosphorylated IKKβ, IKKα, and p65 in VavCre⁺ and Traf6^−/−VavCre⁺ Lin⁻ BM cells. Shown below is the relative expression of the indicated proteins calculated by measuring densitometry of the immunoblots.

(D) Expression and localization of p65 (green) in VavCre⁺ and Traf6^−/−VavCre⁺ LSK cells as determined by indirect immunofluorescence (IF). Shown are representative images from 3 pooled mice per group.

(E) Outline of competitive BM transplantations using IKK^f/fVavCre⁻ or IKK^−/−VavCre⁺ mice.

(F) Summary of donor-derived PB proportions at the indicated time points after competitive transplantation using IKK^f/fVavCre⁻ or IKK^−/−VavCre⁺ BM cells (n = 5 per group). Two independent experiments were performed.

(G) Representative flow cytometric analysis of donor-derived (CD45.2⁺) and competitor-derived (CD45.1⁺) BM and spleen cells from recipient mice after competitive transplantation using IKK^f/fVavCre⁻ or IKK^−/−VavCre⁺ BM cells.

(H) Proportion of donor-derived IKK^f/fVavCre⁻ or IKK^−/−VavCre⁺ BM and spleen cells 8 weeks after competitive BM transplantation (n = 5 per group).

(I) Representative flow cytometric analysis of donor-derived (CD45.2⁺) and competitor-derived (CD45.1⁺) BM LSK cells from recipient mice after competitive transplantation using IKK^f/fVavCre⁻ or IKK^−/−VavCre⁺ BM cells.

(J) Proportion of donor-derived IKK^f/fVavCre⁻ or IKK^−/−VavCre⁺ LSK BM 8 weeks after competitive BM transplantation (n = 5 per group). *p < 0.05. Data are represented as mean ± SEM.