Figure 2: PPARγ binds p65.
From: PPARγ is an E3 ligase that induces the degradation of NFκB/p65
(a) Confocal analysis of PPARγ interaction with p65 in HT29 cells. Arrow heads show PPARγ co-localized with p65. Scale bar, 30 μm. (b) HT29 or MEF cell lysates were subjected to immunoprecipitation and western blot analysis as shown. (c) HEK293T cells were transfected with GST-p65 and HA-PPARγ and subcellular fractions were subjected to GST pull-down (GST-PD) and western blotting. To inhibit protein degradation, cells were treated with 10 μM MG132 for 6 h before cell lysis. (d) Constructs of p65 (upper panel). Cell lysates of HA-PPARγ, GST-RHD or GST-307-551 transfected in HEK293T cells were subjected to GST pull-down and western blotting. Cells were treated with 10 μM MG132 for 6 h before cell lysis. (e) Constructs of PPARγ (upper panel). Cell lysates of p65, GST-PPARγ or GST-PPARγ-ΔLBD transfected in HEK293T cells were subjected to GST pull-down and western blotting. Cells were treated with 10 μM MG132 for 6 h before cell lysis. Data are triplicates from three independent experiments. DAPI, 4′,6-diamidino-2-phenylindole.
