Journal: Oncotarget

Article Title: TNFSF10/TRAIL regulates human T4 effector memory lymphocyte radiosensitivity and predicts radiation-induced acute and subacute dermatitis

doi: 10.18632/oncotarget.7893

Figure Lengend Snippet: A. Dose dependent apoptosis of T4EM lymphocytes monitored by AnnexinV fluorescence intensity 18 hours after irradiation. B. Ranking of 373 individuals according to their T4EM lymphocytes radiosensitivity measured by the level of apoptosis (grey circles). T4EM lymphocytes from four “sensitive” and four “resistant” unrelated individuals (black circles) were selected for micro-array expression analysis. T4EM lymphocytes from fifteen “sensitive” (red), “median” (green) and “resistant” (blue) unrelated individuals were used for qPCR. C. Quantitative PCR analysis of expression level of TRAIL/TNFSF10 in sorted T4EM lymphocytes from 15 “resistant” (R), 15 “median” (M), and 15 “sensitive” unrelated individuals (S). Results are shown as box plots of dCt with respect to the internal reference gene ( RPLP0 ) with median values (red) (*** p < 10 −3 ). D. Multiplexed qPCR analysis of TNFSF10/TRAIL (left panels) , TNFRSF10a/DR4 (middle panels) and TNFRSF10b/DR5 (right panels) in sorted T4EM lymphocytes from 5 “resistant” (R) and 5 “sensitive” (S) unrelated individuals, 2, 4, 6, and 24 hours after irradiation at 2 Gy. Results are shown as box plots with median line (red) (*** p < 10 −3 ) that represent, for each time point post-irradiation (hours), the difference in dCt (ddCt) between non-irradiated sample and the dCt of the test sample.

Article Snippet: Recombinant human soluble TRAIL (rh-sTRAIL; Cat# 375-TL), anti-TRAIL cytotoxicity blocking antibody (CD253; clone RIK-2) and recombinant human soluble DR5 Fc chimera (rh-sDR5/Fc; Cat#631-T2) were from R&D Systems, AnnexinV-FITC from Becton Dickinson, and 1,10-phenanthroline from Sigma-Aldrich.

Techniques: Fluorescence, Irradiation, Microarray, Expressing, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: TNFSF10/TRAIL regulates human T4 effector memory lymphocyte radiosensitivity and predicts radiation-induced acute and subacute dermatitis

doi: 10.18632/oncotarget.7893

Figure Lengend Snippet: A. mTRAIL level, quantified by flow cytometry, on resting T4EM lymphocytes from 4 “sensitive” (S) and 4 “resistant” (R) samples from unrelated individuals. Results are shown as box plots with median line (red) (*** p < 10 −3 ). B. Apoptosis of T4EM lymphocytes monitored by AnnexinV fluorescence intensity of “resistant” (left panel) or “sensitive” (right panel) samples after 18 hours without irradiation (shaded grey), or after 2 Gy irradiation in the absence (“resistant” blue / “sensitive” red) or presence (black) of TRAIL blocking antibody (CD253). C. Percentage of apoptotic T4EM lymphocytes from 4 “sensitive” (S) and 4 “resistant” (R) samples, 18 hours after irradiation at 0, 0.5, 1, and 2 Gy, in the absence (−) or presence (+) of TRAIL blocking antibody (CD253). Results are shown as box plots with median line (red) (* p < 0.05, *** p < 10 −3 ). D. Apoptosis of T4EM lymphocytes monitored by AnnexinV fluorescence intensity of “resistant” (left panel) or “sensitive” (right panel) samples after 18 hours without irradiation (shaded grey), or after 2 Gy irradiation in the absence (“resistant” blue / “sensitive” red) or presence of 0.4 μg/ml rh-sTRAIL (black). E. Percentage of apoptotic T4EM lymphocytes from 4 “sensitive” (S) and 4 “resistant” (R) samples, 18 hours after irradiation at 0, 0.5, and 2 Gy, in the absence (−) or presence (+) of rh-sTRAIL (0.4 μg/ml). Results are shown as box plots with median line (red) (* p < 0.05, *** p < 10 −3 ). F. Percentage of apoptotic T4EM lymphocytes from 4 “sensitive” (S) samples, 18 hours after irradiation at 0 and 2 Gy, in the absence (−) or presence (+) of rh-sDR5/Fc (40 ng/ml). Results are shown as box plots with median line (red) (*** p < 10 −3 ). G. Radiosensitivity of 9 samples from unrelated “sensitive” individuals after 2 Gy irradiation at cell concentrations of 10 5 /ml (1X) and 10 6 /ml (10X) (** p < 10 −2 ).

Article Snippet: Recombinant human soluble TRAIL (rh-sTRAIL; Cat# 375-TL), anti-TRAIL cytotoxicity blocking antibody (CD253; clone RIK-2) and recombinant human soluble DR5 Fc chimera (rh-sDR5/Fc; Cat#631-T2) were from R&D Systems, AnnexinV-FITC from Becton Dickinson, and 1,10-phenanthroline from Sigma-Aldrich.

Techniques: Flow Cytometry, Fluorescence, Irradiation, Blocking Assay