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

Figure 1. CCE9 induces Nur77 expression and apoptosis.

Figure 1

(A) Structure of CCE9. (B) Time-course analysis of Nur77 and PARP cleavage induction by CCE9. HeLa229 cells treated with 10 μM CCE9 for the indicated time were determined by Western blotting using anti-Nur77 antibody. (C) Dose dependent effect of CCE9. HeLa229 cells treated with vehicle or indicated concentration of CCE9 for 3 hr were analyzed for Nur77 expression and PARP cleavage by Western blotting. (D) Time-course analysis of Nur77 expression and apoptosis induction by CCE9 in A549 and HepG2 cells. Cells treated with 10 μM CCE9 for the indicated time were analyzed for Nur77 expression and PARP cleavage by Western blotting. (E) Dose-dependent induction of Nur77 and apoptosis by CCE9 in A549 and HepG2 cells. Cells treated with vehicle or the indicated concentration of CCE9 for 3 hr were analyzed for Nur77 expression and PARP cleavage by Western blotting. (F) RT-PCR analysis of Nur77 mRNA expression in HeLa229 cells. Cells treated with vehicle, TPA (100 ng/ml), or indicated concentration of CCE9 for 3 hr. Nur77 and β-actin mRNA products were simultaneously amplified in the same reaction system, in which β-actin expression level served as an internal control. (G) Caspase-3 activation by CCE9. HeLa229 cells were treated with 10 μM CCE9 for 3 hr, immunostained with antibody recognizing the cleaved caspase-3. Nuclei were visualized by co-staining with DAPI. (H) DAPI staining. HeLa229 cells were treated with CCE9 (10 μM) or vehicle for 3 hr or 6 hr and subjected to DAPI staining. Apoptotic cells were scored and compared between different treatments. *, P<0.01 (VS. control); **, P<0.01 (VS. control). (I) The apoptotic effect of CCE9. HeLa229 cells were treated with vehicle or 10 μM CCE9 for 6 hr and stained with Annexin V/PI. Apoptosis was analyzed by fluorescence-activated cell sorting analysis.