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. 2013 Jul 25;41(18):8515–8525. doi: 10.1093/nar/gkt624

Figure 4.

Figure 4.

The effect of Pin1 inhibition on GR phosphorylation, nuclear translocation and stability. (A) A549 cells were transfected with Pin1 or control siRNA for 48 h. Cells were then treated with 1 or 100 nM for 30 min. Immunoblots were probed for phospho- GR (S211), GR, Pin1 and tubulin. (B) Cells were pre-treated with juglone for 30 min before a 30-min treatment of 100 nM DEX. (C) A549 cells were pre-treated with juglone (30 µM) for 30 min before a 30-min treatment with 100 nM DEX. Cytosol and nuclear fractions were prepared, and subsequent immunoblots were probed for GR and Histone H1. (D) A549 cells were transfected with Pin1 or control siRNA for 48 h before being stimulated with DEX (100 nM) or 4 or 8 h, whole-cell extracts were probed for GR and Pin1. (E) A549 cells were transfected with Pin1 siRNA as described in (D), cycloheximide (50 µg/ml) was added to the cells for 4, 8 and 16 h, subsequent immunoblots were probed for GR, Pin1 and β-actin.