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Fig. 3 | Retrovirology

Fig. 3

From: Hypericin-photodynamic therapy inhibits the growth of adult T-cell leukemia cells through induction of apoptosis and suppression of viral transcription

Fig. 3

Hypericin-PDT induced caspase dependent and independent apoptosis. a Effects of the general caspase inhibitor z-VAD-fmk on hypericin-PDT induced growth inhibition of TL-Om1 cells. Cells were pretreated for 2 h with 10 μM z-VAD-fmk followed by 24 h treatment with 30 ng/mL hypericin. The proliferation of each cell was examined by MTT assay, b protein levels of caspase family proteins hypericin-PDT-treated cells. The expression of Caspase 3, Caspase 7, Caspase 8, and Caspase 9 were analyzed by immunoblotting. ← indicates cleavaged caspase protein, c hypericin-PDT treatment influenced the expression of apoptosis-related proteins. The expression of Bax, Bid, Bcl-2, Bad, and Cyt-C were analyzed by western blot. ← indicates cleavaged Bid protein, d z-VAD-fmk reversed the modulating function of hypericin-PDT on apoptosis-related proteins’ expression. HPB-ATL-T cells were pretreated for 2 h with 10 μM z-VAD-fmk followed by 24 h treatment with hypericin (150 ng/mL)-PDT. The expression of cleaved Caspase 3, cleaved Caspase 9, Bax, and Bcl-2 were analyzed by immunoblotting, e hypericin enhanced Bax promoter activity in TL-Om1 and HPB-ATL-T cells. Cells were transfected with the Bax reporter plasmid with or without hypericin-PDT treatment. Luciferase activity was measured 48 h after transfection, f western blot analysis for the expression of p53, PARP, and GAPDH proteins in hypericin-PDT-treated ATL cells. ← indicates cleavaged PARP protein, g hypericin-PDT activated p53 signaling pathway. Hypericin-PDT-treated HPB-ATL-T cells were cotransfected with p53-Luc and phRL-TK. After 48 h, the cells were harvested and analyzed for luciferase activity, h hypericin-PDT treatment influenced the expression of AIF protein. The expression of AIF and GAPDH were analyzed by western blot. Statistically significant differences are labeled *P < 0.05, **P < 0.01, compared with control group using Student’s t-test

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