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. Author manuscript; available in PMC: 2014 May 20.
Published in final edited form as: Nat Immunol. 2012 Jan 22;13(3):300–307. doi: 10.1038/ni.2210

Figure 5. A positive feedback loop comprised of Foxo1-BLNK-p38 that regulates pre-B cell differentiation.

Figure 5

(a) and (b) Immunoblot analysis of indicated proteins in IRF-4,8−/− pre-B cells after transduction with Foxo1 or Blnk retroviral vectors. Cells were maintained in IL-7 Hi conditions and sorted based on GFP expression after transduction as described in Fig. 1f. MigR, MigR-Foxo1 and MigR-Blnk denote control and Foxo1 or Blnk encoding retroviruses, respectively. (c) Immunoblot analysis of phospho-p38 (p-p38), total p38 (p38), Foxo1 and HPRT in the absence or presence of the p38 inhibitor SB203580 (SB, 0.1 μg/mL) and the indicated IL-7 culture conditions. (d) Foxo1 binding to regulatory elements in the Rag and Blnk loci under the conditions described above. ChIP assay was performed using αFoxo1 antibodies as in Fig. 1c. (e) Immunoblot analysis of Foxo1, Blnk, p-p38, p38 and HPRT upon attenuation of IL-7 signaling. IRF-4,8−/− pre-B cells were cultured in IL-7 Lo conditions and whole cell lysates were prepared and analyzed by immunoblotting at the indicated time points. Data are representative of three independent experiments.