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. 2012 May 24;120(1):155–165. doi: 10.1182/blood-2011-05-357418

Figure 4.

Figure 4

Sox4 overexpression enhances proliferation and self-renewal of myeloid cells. (A) WT and TG mouse bone marrow was transduced with the pMIG-Sox4 overexpressing vector or pMIG retroviral vector, sorted for GFP positivity, and seeded into semisolid MethoCult media. Colony scoring was performed 7 to 14 days after seeding or replating. Sox4 expression enhances self-renewal of cells in in vitro semisolid cultures. Sox4 expression in combination with overexpression of the proto-oncogene CREB enhances both proliferation and replating of mouse bone marrow transductants (ANOVA test P ≤ .001, Tukey analysis P values comparing groups/columns is represented above the data bars; *P < .05, **P < .01, ***P < .001) in 3 separate experiments. (Inset) Immunoblot detecting the expression of sox4, CREB, and actin-loading control of colonies grown on MethoCult. (B) Immunophenotypic characterization of WT and TG mouse bone marrow cells transduced with Sox4 retroviral construct or empty vector, sorted, and grown in liquid media supplemented with cytokines. CREB and sox4 synergize to enhance the proliferation or survival of Gr+Mac+ myeloid cells. (C) Graphical representation of panel B (1-way ANOVA test (P < .001). Tukey analysis shows a significant difference (P ≤ .05) in Gr-1+Mac-1+ percentages between WT vector vs TG vector, TG vector vs TG sox4, and WT sox4 vs TG sox4. Analysis was based on 3 independent experiments performed in triplicate.