Fig. 5
From: SENP3 maintains the stability and function of regulatory T cells via BACH2 deSUMOylation
SENP3 mediates BACH2 deSUMOylation in Treg cells. a SENP3-BACH2 co-immunoprecipitation (co-IP) assays using HEK293 cells transfected with the indicated expression vectors. b Lysates from Senp3+/+Foxp3-Cre and Senp3fl/flFoxp3-Cre splenic Treg cells stimulated with anti-CD3 and anti-CD28 antibodies for 24 h were subjected to IP using anti-BACH2 antibody or control Ig (Ctrl); BACH2 and BACH2-associated SENP3 were detected by immunoblotting (IB). c ChIP assay to evaluate the binding of BACH2 to the regulatory regions of Il4, Il5, and Ccr4 loci in Senp3fl/flFoxp3-Cre and Senp3+/+Foxp3-Cre splenic Treg cells stimulated with anti-CD3 and anti-CD28 antibodies for 24 h. d, e BACH2 SUMOylation assays using HEK293 cells transfected with the indicated expression vectors. f BACH2 SUMOylation assays using Senp3+/+Foxp3-Cre and Senp3fl/flFoxp3-Cre Treg cells stimulated with anti-CD3 and anti-CD28 antibodies for 0, 4 or 8 h. g Flag-tagged mouse BACH2 or its mutant variants were individually transfected into HEK293T cells for BACH2 SUMOylation assays. h Sequence alignment of the K275 and K579 SUMOylation sites of BACH2 in different species and conserved residues (red) are shown. i Flag-tagged mouse BACH2 (WT) or K275R and K579R mutant BACH2 (2KR) were individually transfected into HEK293T cells for BACH2 SUMOylation assays. All data shown are representative of three independent experiments. Error bars are the mean ± SEM values (c). Two-tailed unpaired Student’s t tests were performed (c). *P < 0.05
