IMMUNOLOGY Correction for “14-3-3σ regulates B-cell homeostasis through stabilization of FOXO1,” by Yu-Wen Su, Zhenyue Hao, Atsushi Hirao, Kazuo Yamamoto, Wen-Jye Lin, Ashley Young, Gordon S. Duncan, Hiroki Yoshida, Andrew Wakeham, Philipp A. Lang, Kiichi Murakami, Heiko Hermeking, Bert Vogelstein, Pamela Ohashi, and Tak W. Mak, which appeared in issue 4, January 25, 2011, of Proc Natl Acad Sci USA (108:1555–1560; first published January 4, 2011; 10.1073/pnas.1017729108).
The authors note that Fig. 5 appeared incorrectly. The figure has been revised to indicate where gels were rearranged and where extraneous data has been removed. The corrected figure and its legend appear below.
Fig. 5.
FOXO functions. (A) Increased FOXO target gene (Bim) mRNA. Splenic B cells were left untreated (non) or stimulated for 2 or 6 h with 10 μg/mL anti-IgM or 10 μg/mL anti-IgM plus 1 μg/mL CD40L (IgM/CD40). Bim mRNA was determined by qRT-PCR. Data are Bim mRNA relative to 18S RNA. ΔΔCt values were normalized to WT values in untreated controls (fold change, 1). *P < 0.05. (B) Altered proteins. Western blot of FOXO1, p27, Bim, and β-actin proteins in resting WT and KO B cells. (C) Normal FOXO1 mRNA. qRT-PCR of FOXO1 mRNA in resting WT and KO B cells. (D) Restoration of FOXO1 protein by proteasome inhibition. Purified WT and KO B cells were left untreated (–) or treated with MG132 (20 μM) for 1 h at 37 °C. Nuclear (N) and cytosolic (C) fractions of cells were immunoblotted to detect FOXO1 protein, P-S6 ribosomal protein (Ser240/244; p-rpS6), and total S6 ribosomal protein (rpS6; loading control for cytosol). (E) FOXO1 interacts directly with 14-3-3σ protein under overexpression conditions. 293T cells were transiently transfected with plasmid encoding His-tagged 14-3-3σ, without (–) or with (+) plasmid encoding Flag-tagged FOXO1. At 2 d after transfection, total lysates were immunoprecipitated with beads conjugated to anti-His (IP: His) or anti-Flag (IP: Flag) and immunoblotted to detect FOXO1 and 14-3-3σ. (F) FOXO1 interacts directly with endogenous 14-3-3σ in the cytosol. Ramos B cells were left untreated (0) or stimulated with anti-human IgM for 5 or 30 min. Nuclear (N) and cytosolic (C) fractions of cells were immunoprecipitated with anti–14-3-3σ or anti-FOXO1 and immunoblotted to detect 14-3-3σ and FOXO1. (G) Altered signaling downstream of BCR engagement. Purified WT and KO B cells were left untreated (0) or stimulated with 10 μg/mL anti-IgM for the indicated times. Lysates were immunoblotted to detect P-Erk1/2, total Erk1/2, P-JNK1/2, P-Akt (S473), and total Akt.