FIGURE 2.

Proline-rich region of CBL-Y371H mutant is essential for its gain-of-function phenotype in HSPCs. A, CBL/CBL-B DKO mouse HSPCs were isolated, retrovirally infected with the indicated CBL constructs, and infected (GFP+) cells FACS-sorted as in Fig. 1, followed by analysis of SCF-induced proliferation (1 ng/ml) using the Cell TiterGlo assay. B–D, CBL-null mouse HSPCs were retrovirally infected with the indicated CBL constructs, and GFP+ cells were assessed in three replicates for cytokine-independent (B) or SCF-stimulated proliferation (C) (Cell TiterGlo), or for SCF-stimulated colony forming ability (D). Proliferation based on luminescence arbitrary unit in A–C was normalized to the vector-only control. Shown is pooled data of three independent experiments demonstrated as percentage to vector-only control (mean ± S.E.).