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. 2012 Jun 25;2:23. doi: 10.1186/2045-3701-2-23

Figure 5.

Figure 5

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.