Figure 5
From: PDLIM2 restricts Th1 and Th17 differentiation and prevents autoimmune disease
Ubiquitination and proteasomal degradation of STAT3 by PDLIM2. A) Physical interaction between PDLIM2 and STAT3. Nuclear extracts of 293 cells transfected with HA-STAT3 alone or together with Myc-PDLIM2 were subjected to immunoprecipitation (IP) using Myc antibody and immunoblotting (IB) using HA antibody. The expression levels of HA-STAT3 and Myc-PDLIM2 were examined by IB. B) Polyubiquitination of STAT3 by PDLIM2. 293 cells were transfected with HA-STAT3 plus Flag-ubiquitin in the presence or absence of Myc-PDLIM2, followed by nuclear fractionation. The nuclear extracts were subjected to IP using HA antibody and IB using Flag antibody. The expression levels of HA-STAT3 and Myc-PDLIM2 were examined by IB. C) Proteasomal degradation of STAT3 by PDLIM2. 293 cells transfected with HA-STAT3 alone or together with Myc-PDLIM2 were cycloheximide (CHX)-chased for the indicated time, followed by nuclear extractions and IB using HA or Myc antibody. In lanes 3 and 6, the cells were chased in the presence of 10 μM MG132.