Fig. 8

Bcl2l11 knockdown and Irf8 overexpression simultaneously restored cDC1 generation from TRIM33-deficient BM Lin^– precursors. A Scheme of the Lin^– cell in vivo reconstitution experiment. Lin^– cells were isolated from Trim33^fl/fl (WT) or Trim33^fl/fl Itgax-Cre (cKO) BM, expanded with Flt3L and SCF, transduced with shRNA constructs, and i.v. injected into recipients. B Bcl2l11 knockdown efficiency in transduced mAmetrine⁺ WT and cKO Lin^– cells. C–E Flow cytometry analysis of splenic DC populations in the indicated Lin^– reconstituted mice. Pregate: viable, CD45.2⁺mAmetrine⁺ cells. The numbers on the plots indicate percentages. D Numbers of CD45.2⁺mAmetrine⁺ splenic DCs per animal from the indicated mice. E Fold increase in the number of each DC population of the indicated genotypes after Bcl2l11 knockdown. N = 6–10. F Irf8 overexpression and Bcl2l11 knockdown efficiency in BM Lin^– cells. BM Lin^– cells of the indicated genotypes were cotransduced with overexpression and shRNA vectors as previously described. G, H Flow cytometry analysis of DCs generated from Lin^– BM cells transduced with the indicated construct pairs by Flt3L culture. Pregate: viable, CD45.2⁺GFP⁺mAmetrine⁺ cells. H Numbers of DCs generated per 1\(\times\)10⁴ input Lin^– cells. N = 3. Bars represent the means ± SEMs. The data are representative of 2–3 independent experiments. The data were analyzed using unpaired two-tailed multiple t tests (D) or two-way ANOVA followed by Bonferroni’s multiple comparison test (H). Ns: nonsignificant, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001