Supplementary Figure 5: TAZ blocks acetylation of Foxp3 and promotes its degradation.

(a) Real-time quantitative PCR (RT-qPCR) analysis of the expression levels of Foxp3 in CD4⁺ naïve T cells cultured with anti-CD3/CD28 antibodies in the presence of TGF-β (2.5 ng/ml) or TGF-β plus IL-6 (30 ng/ml) for 1 and 2 days. Data were assessed with Student’s t-test and are represented as mean ± s.d. ns, not significant. (b) Immunoassays of 293T cells expressing various combinations (above lanes) of HA-tagged Tip60, and full-length (FL) or the truncated fragment of Flag-TAZ as indicated; immunoprecipitation with anti-Flag and analysis by immunoblot with the indicated antibodies; below, immunoblot analysis of total cell lysates (TCL) without immunoprecipitation. (c) Immunoassays (as in b) of 293T cells expressing various combinations (above lanes) of GFP-tagged TAZ, full-length (FL) or the truncated fragment of Flag-Tip60 as indicated. (d) Immunoassays (as in b) of 293T cells expressing various combinations (above lanes) of GFP-tagged Foxp3, and full-length (FL) or the truncated fragment of Flag-Tip60 as indicated. (e) Immunoassays (as in b) of 293T cells expressing various combinations (above lanes) of Myc-tagged Tip60, Flag-Foxp3 and increasing doses of HA-tagged TAZ. (f) SIM of HeLa cells transfected with NLS/Flag-tagged TAZ (purple), HA-tagged Tip60 (red) or Flag-Foxp3 (green); Scale bars, 20 μm. The data are representative of three independent experiments with similar results (a-f), or with ~50 cells (f).