Figure 1.

Cytokine-Stimulated GADD45G Expression Induces and Accelerates Differentiation in LT-HSCs

(A and B) Quantitative RT-PCR of Gadd45 genes in LT-HSCs (A) and MPPs (B) stimulated with cytokines. n = 3 experiments. Relative expression normalized to B2m.

(C–F) (C) Experimental scheme of (D)–(F). (D) Representative FACS blots of cultured LT-HSCs, gated for GADD45G and control-transduced cells (VENUS⁺).

(E and F) Quantification of FACS results after 5, 8, and 10 days of culture. Transduced cells (VENUS⁺) were gated for immature HSPCs, GMP-like cells and mature GM cells. n = 3 experiments.

(G) Colony formation assay of 200 LT-HSCs transduced with GADD45G and control and replating of 2,000 cells after 10 days. n = 3 experiments.

(H) Experimental scheme of (I) and (J).

(I) Generation times of transduced LT-HSCs (generation 1) and their progeny (TOMATO⁺) in subsequent generations.

(J) Interval of transgene expression (TOMATO⁺) and onset of CD16/32 expression (GMP-like stage). Data extracted from 29 (G45G) and eight pedigrees (control).

(K) Differentiation kinetics of double-transduced LT-HSCs (NA10, NUP98-HOXA10 homeodomain and GADD45G) gated on immature HSPCs by FACS. n = 3 experiments.

Data are represented as mean ± SD. ^∗p < 0.05, ^∗∗p < 0.01. ns, not significant. See also Figure S1.