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. 2017 Jul 13;8(41):69731–69745. doi: 10.18632/oncotarget.19227

Figure 2. Role of Nur77 in CCE9 induced apoptosis.

Figure 2

(A-B) Transfection of Nur77 siRNA inhibits apoptosis induction by CCE9. HeLa229 cells transfected with control or Nur77 siRNA were treated with CCE9 (10 μM) for 3 hr, and analyzed for PARP cleavage by Western blotting (A), caspase-3 activation by immunostaining (B). (C) Transfection of Nur77 promotes apoptosis induction by CCE9. HeLa229 cells transfected with GFP-Nur77 expression vector were treated with 10 μM of CCE9 for 3 hr. Subcellular localization of GFP-Nur77 was examined by confocal microscopy. (D) Nur77 mediates the apoptotic effect of CCE9. HeLa229 cells were transfected with Nur77 siRNA and GFP-Nur77and subjected to CCE9 treatment (10 μM) or vehicle in serum-free medium for 3 hr. Apoptotic cells examined by DAPI staining were compared between different cells. *, P < 0.01 (vs. control); **, P < 0.01 (vs. control); ***, P < 0.01 (vs. control siRNA with CCE9 treatment); #, P>0.05 (vs. control). (E) The apoptotic effect of CCE9 is impaired in Nur77 knockdown and Nur77 overexpressed cells. HeLa229 cells transfected with Nur77 siRNA and HeLa229 cells transfected with GFP-Nur77 were treated with vehicle or 10 μM of CCE9 for 6 hr and stained with Annexin V/PI. Apoptosis was analyzed by fluorescence-activated cell sorting analysis.