Fig. 4.

Daxx regulates PPARγ nucleus transportation. a Confocal picture showed the colocalization of PPARγ (Green) and Daxx (Red) in the MEF of WT mice; scale bar 20 μm. b HEK293 cells were co-transfected with GFP-PPARγ and Myc-Daxx. Immunoprecipitation was performed with the anti-Myc antibody. Immunoblotting was performed with anti-Myc or anti-GFP antibodies. c The truncated Daxx plasmid construction. d HEK293 cells were co-transfected with HA-PPARγ and Myc-tagged Daxx functional domain deletions. Immunoprecipitation was performed with the anti-Myc antibody. Immunoblotting was performed with anti-Myc or anti-HA antibody. e, f Nuclear and cytoplasmic proteins were isolated from the HA-PPARγ and siDaxx-transfected HEK293 cells, and determined by SDS-PAGE. N = 3, unpaired two-tailed Student’s t test, **P < 0.01. g, h HEK293 cells were transfected into GFP-tagged full-length PPARγ by accompany with siNC or siPdcd4 for 24 h, and localizations of GFP were observed by immunofluorescence, and the mean fluorescence intensity of GFP was quantified from n > 30 cells; scale bar 20 μm. Unpaired two-tailed Student’s t test, **P < 0.01. i, j Nuclear and cytoplasmic proteins were isolated from the HA-PPARγ and Daxx-transfected HEK293 cells, and determined by SDS-PAGE. N = 3, unpaired two-tailed Student’s t test, *P < 0.05. k, l HEK293 cells were transfected into GFP-tagged full-length PPARγ by accompany with Myc-Daxx for 24 h, and localizations of GFP were observed by immunofluorescence, and the mean fluorescence intensity of GFP was quantified from n > 30 cells; scale bar 20 μm. Unpaired two-tailed Student’s t test, **P < 0.01. The figures represent three independent experiments that yield similar result