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. 2012 Dec 20;7(12):e52572. doi: 10.1371/journal.pone.0052572

Figure 4. Green tea polyphenols-mediated apoptosis induced by death receptor pathway in prostate cancer LNCaPshV and LNCaPshp53 cells.

Figure 4

[A] Cells were treated with 40 µg/ml concentration of GTP for 1, 5, 10, 15, 30 and 60 min and Western blotting was performed for SAPK/JNK, p-JNK, FAS, FADD, p-FADD, BID and t-BID proteins. A typical actin blot demonstrates internal loading control. [B] Cells were treated with 40 µg/ml concentration of GTP for 3, 6, 9, 12, 24 and 48 h and Western blotting was performed for cleaved PARP, procaspase-8, cleaved caspase-8, c-IAP and XIAP proteins. A typical actin blot demonstrates internal loading control. [C] Cells were treated with 20 µM concentration of JNK inhibitor SP600125 for 8 h and with 40 µg/ml GTP for 16 h alone, or SP600125 for 8 h followed by GTP treatment in combination followed by Western blotting for JNK, p-JNK, FAS and p-FADD proteins. The expression of native JNK protein was considered as loading control. [D] Cell death measurement was performed by photometric enzyme immunoassay Cell Death Detection ELISA kit. The bars represent mean±SD of at least two independent experiments each performed in duplicate, **p<0.001 represents significant differences as compared to control group. [E] Light microscopic images of LNCaPshV and LNCaPshp53 cells treated with SP600125 and GTP alone or in combination. The details are described in the materials and methods section.