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. 2013 Oct 24;14(11):21215–21226. doi: 10.3390/ijms141121215

Figure 4.

Figure 4

Kae-induced apoptosis in EJ cells involves PTEN. (A) Kae upregulates PTEN expression in a time-dependent manner. EJ cells were treated with 40 μM Kae for the indicated times. Levels of PTEN and Akt were detected by Western blotting. Data are expressed as the mean ± SD from three independent experiments (**p < 0.01 vs. DMSO-only group); (B) siRNA (Si A and Si B) inhibits PTEN expression; control siRNA was used as the negative control (NC) (*p < 0.05, **p < 0.01 vs. NC); (C) PTEN siRNA inhibits Kae-induced apoptosis. PTEN siRNA-transfected EJ cells treated with 40 μM Kae for 24 h resulted in significantly decreased apoptosis, compared with NC siRNA-transfected cells treated with 40 μM Kae. Data represent the mean ± SD of three independent experiments (**p < 0.01 vs. NC + Kae group); (D) PTEN siRNA attenuates the growth-inhibiting effects of Kae. PTEN siRNA- or NC siRNA-transfected EJ cells were treated with 40 μM Kae for 24 h; cell viability was determined by the CCK-8 assay. Data shown are the mean ± SD of three independent experiments; (E) PTEN siRNA blocks Kae-induced cleavage of caspase-3. Western blots of cleaved caspase-3 in PTEN siRNA- or NC siRNA-transfected cells treated with 40 μM Kae for 24 h (**p < 0.01 vs. NC + Kae group); (F) LY294002 enhances Kae-inhibition of EJ cell growth. EJ cells were treated with 40 μM Kae, 20 μM LY294002 or both for 24 h. Cell viability was determined by the CCK-8 assay. Data shown are the mean ± SD of three independent experiments; (G) LY294002 enhances Kae-induced cleavage of caspase-3. EJ cells were treated with 40 μM Kae, 20 μM LY294002 or both for 24 h. Western blots of cleaved caspase-3 were performed.