Journal: PLoS ONE

Article Title: Ionizing Radiation Potentiates Dihydroartemisinin-Induced Apoptosis of A549 Cells via a Caspase-8-Dependent Pathway

doi: 10.1371/journal.pone.0059827

Figure Lengend Snippet: (A) DHA-induced concentration-dependent reduction of cell viability assessed by CCK-8 assay. Cells were incubated with various concentrations of DHA (0, 5, 10, 20, 30 µg/ml) for 48 h, *P <0.05, compared with control; **P <0.01, compared with control. (B) DHA-induced time-dependent reduction of cell viability by CCK-8 assay. Cells were incubated with 20 µg/ml of DHA for different time (0, 12, 24, 36, 48 h). **P <0.01, compared with control. (C) DHA-induced nuclear condensation of cells stained by Hoechst 33258 after treatment for 24 h. Images were recorded using a digital camera with 1280×1280 pixels resolution. Magnification 400. (D) DHA-induced Sub-G 1 cell cycle arrest. Cells were cultured with 20 µg/ml of DHA for 0, 12, 24 or 48 h and then stained with 5 µg/ml of PI before being analyzed by FCM. (E) DHA-induced apoptosis assessed by FCM analysis. Cells were treated with 20 µg/ml of DHA for 0, 24 and 48 h, and then analyzed by FCM after staining with Annexin V-FITC/PI.

Article Snippet: Cell apoptosis detection was also performed by flow cytometry (FCM) analysis using Annexin V-FITC/PI apoptosis detection kit (Bender Medsystems, Vienna, Austria) as previously described, and for each FCM analysis 10,000 events were recorded.

Techniques: Concentration Assay, CCK-8 Assay, Incubation, Control, Staining, Cell Culture

Journal: PLoS ONE

Article Title: Ionizing Radiation Potentiates Dihydroartemisinin-Induced Apoptosis of A549 Cells via a Caspase-8-Dependent Pathway

doi: 10.1371/journal.pone.0059827

Figure Lengend Snippet: (A) DHA induced activation of caspase-8 and -9 assessed by fluorometric assay. Cells were treated with DHA for 36 h. ** P <0.01, compared with control. (B) DHA induced caspase-8- and -9-dependent caspase-3 activation by fluorometric assay. Cells were treated with DHA for 48 h in the presence or absence of zIETD-fmk and zLEHD-fmk, respectively. **P <0.01, compared with control; ## P <0.01, compared with DHA treatment alone. (C) DHA induced caspase-8- and -9-dependent cytotoxicity assessed by CCK-8 assay. Cells were treated with DHA for 24 and 48 h in the presence or absence of zIETD-fmk and zLEHD-fmk, respectively. *8 P<0.01 , compared with control; $ P<0.05 , $$ P<0.01 and & P <0.05, compared with DHA treatment alone. (D) DHA ROS-mediated apoptosis assessed by FCM. ** P<0.01, compared with control; ## P<0.01 compared with DHA alone. (E) DHA induced ROS-dependent caspase-8 activation. ** P<0.01 , compared with control; ## P<0.01 , compared with DHA treatment alone. (F) DHA induced ROS- and caspase-8-dependnent loss of Δψ m determined by FCM analysis. **P <0.01, compared with control; ## P<0.01 , compared with DHA treatment alone. (G) DHA induced caspase-8-dependent caspase-9 activation. * P<0.05 and ** P<0.01 , compared with control; # P<0.05 , compared with DHA treatment alone. (H) Typical fluorescence images of Bid translocation to mitochondria inside single living cell after DHA treatment for 36 h. Control cells show the uniform distribution of Bid, while DHA-treated cells show the co-localization between Bid and mitochondria. Scale Bar: 5 µm.

Article Snippet: Cell apoptosis detection was also performed by flow cytometry (FCM) analysis using Annexin V-FITC/PI apoptosis detection kit (Bender Medsystems, Vienna, Austria) as previously described, and for each FCM analysis 10,000 events were recorded.

Techniques: Activation Assay, Control, CCK-8 Assay, Fluorescence, Translocation Assay

Journal: PLoS ONE

Article Title: Ionizing Radiation Potentiates Dihydroartemisinin-Induced Apoptosis of A549 Cells via a Caspase-8-Dependent Pathway

doi: 10.1371/journal.pone.0059827

Figure Lengend Snippet: (A) FCM analysis of cells cycle after low-dose IR treatment for 24 h and 36 h in the presence or absence of DHA. (B and C): IR potentiated the DHA-induced G 2 /M arrest at 24 h (B) and apoptosis at 36 h (C) analyzed by FCM. **P <0.01, compared with treatment with control; ## P <0.01, compared with DHA treatment alone, $$ P <0.01 compared with 2 Gy IR treatment; && P <0.01, compared with 4 Gy IR treatment. Cells treated with different doses of IR were cultured with 20 µg/ml of DHA for indicated time and then stained with 5 µg/ml of PI before being analyzed by FCM.

Article Snippet: Cell apoptosis detection was also performed by flow cytometry (FCM) analysis using Annexin V-FITC/PI apoptosis detection kit (Bender Medsystems, Vienna, Austria) as previously described, and for each FCM analysis 10,000 events were recorded.

Techniques: Control, Cell Culture, Staining