Fig. 6
From: SENP3 maintains the stability and function of regulatory T cells via BACH2 deSUMOylation
BACH2 deSUMOylation regulates its nuclear export in Treg cells. a IB analysis of the indicated proteins in whole (WL), nuclear (NE), and cytoplasmic (CE) extracts of Senp3+/+Foxp3-Cre and Senp3fl/flFoxp3-Cre Treg cells stimulated with anti-CD3 and anti-CD28 antibodies for 0, 4 or 8 h. bIB analysis of the indicated proteins in anti-CD3 and anti-CD28 antibody-stimulated naive CD4+ T cells from Bach2fl/flCd4-Cre mice transduced with the indicated lentiviral vector encoding Flag-WT BACH2 or Flag-mutant BACH2 under Treg cell conditions. c IB analysis of the indicated proteins in different lentivirus-transduced BACH2-deficient CD4+ T cells under Treg cell conditions treated with 10 ng/ml of leptomycin B for 4 h. d IB analysis of Senp3+/+Foxp3-Cre and Senp3fl/flFoxp3-Cre splenic Treg cells stimulated with anti-CD3 and anti-CD28 antibodies in the presence of leptomycin B for 4 h. e Splenic Treg cells (CD4+CD25+GFP+) from Rag1−/− mice reconstituted (for 8 weeks) with Bach2+/+Cd4-Cre or Bach2fl/flCd4-Cre bone marrow (BM) cells transduced with empty vector (EV) or vector expressing WT or 2KR BACH2 were stimulated with anti-CD3 and anti-CD28 antibodies for 24 h and subjected to flow cytometric analysis of Foxp3 expression. f Splenic Treg cells from Rag1−/− mice reconstituted with Senp3+/+Cd4-Cre or Senp3fl/flCd4-Cre BM cells transduced with EV, WT or 2KR BACH2 were stimulated with anti-CD3 and anti-CD28 antibodies for 24 h and subjected to flow cytometric analysis of Foxp3 expression. Numbers above graphs indicate MFI of Foxp3 (e, f). Data shown are representative of three independent experiments. Error bars are the mean ± SEM values (e, f). Two-tailed unpaired Student’s t tests were performed (e, f). *P < 0.05
