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. 2017 Dec 4;214(12):3543–3552. doi: 10.1084/jem.20170468

Figure 1.

Figure 1.

Lithium treatment induces AID Serine 38 phosphorylation. (A) Anti-pS38 and anti-AID immunoblot of Flag-AID purified with anti-Flag antibody from lysates of 3-d LPS- and IL-4–stimulated AIDF/F B cells treated with indicated concentration of lithium acetate (Li+) for indicated time before harvest. (B) Anti-pS38, anti-AID, and anti–α-tubulin immunoblot of cell lysates from WT B cells treated with 10 mM LiCl (Li+) or NaCl (Na+) as a control for the indicated time. (C) Anti-pS38 and anti-AID immunoblot (light and dark exposures) of AID immunoprecipitated from lysates of CH12 cells cultured with LPS (LIT) or CD40 (CIT) together with IL-4, and TGF-β and treated with 10 mM LiCl or NaCl for 6 h. (D) Representative anti-pS38 and anti-AID immunoblot of AID purified with anti-AID antibody from lysates of WT B cell lysates cultured for 3 d in LPS and IL-4. Cells were treated with 2 µM CHIR99021 or 25 mM LiCl (Li+) for 6 h before harvest. (E) Anti-pS38 and anti-AID immunoblot of Flag-AID purified with anti-FLAG antibody from cultured AIDF/F B cell lysates. Concentration and time of CHIR99021 treatment are indicated. (F) Immunoblot of anti-pS38 and anti-AID (light and dark exposures) of Flag-AID purified with anti-Flag antibody from cell lysates. WT or AIDF/F cultured B cells were treated with 10 µg/ml cycloheximide (CHX) 1 h before addition of 10 mM LiCl for 6 h. All panels representative of n = 3 independent experiments except F, n = 2.