Fig. 7 |. TAZ compartmentalizes TeAD and other transcriptional factors to the nuclear puncta.

a, Myc–TEAD4 was cotransfected into MCF-10A cells together with GFP vector or WT or mutant GFP–TAZ as indicated. TAZ and TEAD4 localization was monitored by GFp fluorescence and using immunofluorescence staining with anti-Myc antibodies (red), respectively. Scale bar, 10 μm. Insets, magnification by 2.96, 2.75, 2.75 and 2.75 times, respectively. b, In vitro droplet-formation assay. mCherry–TEAD4 (50 μM) either alone or mixed together with 50 μM WT GFp–TAZ or ΔWW+ΔCC was analysed using a droplet-formation assay under the same conditions as described in Fig. 3c. Scale bar, 10 μm. c, The ability of HA-tagged WT or mutant TAZ to interact with Flag–TEAD4 was examined using a co-immunoprecipitation assay with anti-Flag antibodies in the immunoprecipitation (Ip), followed by western blotting with anti-HA antibodies (top). The abundance of these proteins in the cell lysates was assessed using western blotting (bottom). d, Serum-starved MCF-10A cells transfected with Myc–TEAD4 and GFP–TAZ were treated with 10% FBS for 1 h and processed for immunofluorescence staining using anti-Myc antibodies (red). Scale bar, 10 μm. Inset, magnification by 3.02 times. e, Colocalization of BrD4, MED1 or CDK9 with GFp–TAZ in the nuclear puncta in MCF-10A cells. Localization of endogenous BrD4, MED1 and CDK9 was detected by indirect immunofluorescence (red). Scale bars, 10 μm. Insets, magnification by 2.85 times. f, Colocalization of active rNA pol II with Flag–TAZ in the nuclear puncta in MCF-10A cells was detected using immunofluorescence staining with antibodies targeting either the active rNA pol II, which is phosphorylated at Ser 5 (S5p) or Ser 2 (S2p) in its CTD (red), or Flag (green). The images were captured using super-resolution structured illumination microscopy. Colocalization (yellow) was analysed using Imaris. Scale bar, 2 μm. g, Localization of active or repressive histone marks in MCF-10A cells expressing GFp–TAZ was analysed using immunofluorescence staining with anti-H3K4me3 or anti-H3K9me3 antibodies (red). The images were captured using super-resolution structured illumination microscopy. Colocalization (yellow) was analysed using Imaris. Scale bar, 2 μm. The experiments in a–g were repeated independently three times with similar results. Source data are available online.