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Molecules 2011, 16 1396 Figure 7. Effect of TαPcZn-PDT on Bcl-2 and Fas respectively in TαPcZn-PDT-induced apoptosis of Bel-7402 cells analyzed by Immunoblot assay. Expression of Bcl-2 and Fas in: Lane 1, control Bel-7402 cells treated with 0.1% DMSO; Lane 2, Bel-7402 cells treated with red-light irradiation (53.7 J/cm2); Lane 3, Bel-7402 cells treated with 54 μM TαPcZn; and Lane 4~6, Bel-7402 cells treated with 18, 36, 54 μM TαPcZn in the presence of red- light irradiation(53.7 J/cm2), respectively. 3. Experimental Section 3.1. Materials Anti-Bcl-2 and anti-Fas antibodies were purchased from Santa Cruz Biotechnology (CA, USA). Annexin-V-FLUOS Staining Kit was purchased from Roche (Basel, Switzerland). Western blotting kit was purchased from Invitrogen (CA, USA). DMSO, MTT, PI, and RNAase were purchased from Sigma (St. Louis, MO, USA). RPMI-1640 medium was purchased from Gibco (CA, USA). Fetal bovine serum was purchased from PAA (Coelbe, Germany). Penicillin was purchased from Harbin Pharmaceutical Group (Heilongjiang, China). Streptomycin was purchased from Dalian Merro Pharmaceutical (Liaoning, China). Polyvinylidene difluoride membrane (PVDF) membrane was purchased from Amersham Pharmacia Biotech (Piscataway, NJ, USA). TαPcZn was synthesized as described in our previous report [33]. The TαPcZn stock solution was prepared in DMSO and stored at 4 °C in the dark. When used, the stock solution was appropriately diluted to obtain the desired concentration with a final DMSO concentration of 0.1%. All other chemicals and reagents were of analytic grade. 3.2. Ultraviolet-visible absorption spectrum assay UV-vis absorption spectrum of TαPcZn in DMSO/water (1:3, v/v) mixtures was assayed by UV- 2550 Spectrophotometer (Shimadzu, Kyoto, Japan). 3.3. Cell culture Hepatocellular carcinoma Bel-7402 cells and HDFs were obtained from the Institute of Zoology, Chinese Academy of Science, China and cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and 100 U/mL penicillin-100 mg/L streptomycin in a humidified atmosphere of 5% CO2 at 37 °C.PDF Image | Zinc-Mediated Photodynamic Therapy Inhibits the Proliferation
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