Journal: Human Gene Therapy

Article Title: CD46-Mediated Transduction of a Species D Adenovirus Vaccine Improves Mucosal Vaccine Efficacy

doi: 10.1089/hum.2013.215

Figure Lengend Snippet: Species C and D Ad tissue tropism. Species C and D viruses expressing GFPLuc were used to infect human lung carcinoma cells (A549) (B), human T lymphocyte (Jurkat) (C), mouse macrophages (RAW 264.7) (D), and human dendritic cells (CD14+) (E). Representative images are shown at 400× magnification (A). The cells were lysed at 24 hr postinfection using 5× passive lysis buffer. Luciferase assay reagent was added and luminescence was measured using a Beckman Coulter DTX 880. Luminescence values represent the luciferase activity of the entire well of a 6-well plate. Ad, adenovirus. Color images available online at www.liebertpub.com/hum

Article Snippet: The plate was shaken on an orbital shaker and luciferase activity was measured using a Beckman Coulter DTX 880.

Techniques: Expressing, Lysis, Luciferase, Activity Assay

Journal: International Journal of Nanomedicine

Article Title: Immunotoxicity of titanium dioxide nanoparticles via simultaneous induction of apoptosis and multiple toll-like receptors signaling through ROS-dependent SAPK/JNK and p38 MAPK activation

doi: 10.2147/IJN.S176087

Figure Lengend Snippet: Effects of TiO 2 NPs on the antioxidant defense protein of RAW 264.7 cells. Notes: ( A ) SOD enzyme activity; ( B ) quantitative analysis of relative fold change in Nrf2 protein levels; ( C ) Western blot image of Nrf2 protein. Cells were incubated with indicated concentrations of TiO 2 NPs for 6 hours. Briefly, 20 µg/mL of protein sample was taken and the SOD activity (inhibition rate%) was estimated by BioVision SOD Activity Assay Kit using a multimode microplate reader (DTX-880, Bechman Counter Inc.) at 450 nm. In Western blot, 20 µg/well of protein samples were loaded on 10% SDS-PAGE and transferred to PVDF membrane. Protein blot signals were detected under UVP Biospectrum-600 imaging system (Thermo Fisher Scientific). Protein intensity was quantified using ImageJ. β-Actin antibody was used as control to normalize the data for Western blot. Data are presented as the mean ± standard error of mean; * P <0.05, ** P <0.01, and *** P <0.001 indicate significant differences when tested with ANOVA. Tukey’s test was used for post hoc tests. Abbreviations: TiO 2 , titanium dioxide; NPs, nanoparticles; Nrf2, nuclear factor erythroid 2-related factor 2; SOD, super oxide dismutase; PVDF, polyvinylidene fluoride.

Article Snippet: After-ward, 10 µL/well CCK-8 solutions were added, incubated for 1 hour and absorbance was measured at 450 nm using DTX-880 multimode microplate reader (Beckman Coulter, Inc., Brea, CA, USA).

Techniques: Activity Assay, Western Blot, Incubation, Inhibition, SDS Page, Imaging

Journal: International Journal of Nanomedicine

Article Title: Immunotoxicity of titanium dioxide nanoparticles via simultaneous induction of apoptosis and multiple toll-like receptors signaling through ROS-dependent SAPK/JNK and p38 MAPK activation

doi: 10.2147/IJN.S176087

Figure Lengend Snippet: TiO 2 SA20(−) NPs induced cytotoxicity in murine macrophages. Notes: ( A ) Inverted micrograph of TiO 2 SA20(−) NPs induced cytotoxicity in RAW 264.7 cells (bar, 50 µm; magnification, ×20). ( B ) Effects of TiO 2 NPs on the viability of macrophage Raw 264.7 cells. Cells were incubated with indicated concentrations of TiO 2 SA20(−) NPs for 24 hours. After NPs exposure, live macrophage cells were analyzed to detect cytotoxicity under digital inverted microscope (EVOS ® XL, Life Technologies Co., Ltd.). Later, cell viability was assessed using CCK-8 solutions (Dojindo Lab.) for 1 hour and absorbance was measured at 450 nm using DTX-880 multimode microplate reader (Beckman Coulter, Inc.). Untreated cells were considered as control in this experiment. Data are presented as the mean ± standard error of mean; *** P <0.001 indicates significant difference when tested with ANOVA. Tukey’s test was used for post hoc tests. Abbreviations: TiO 2 , titanium dioxide; NPs, nanoparticles; CCK-8, cell counting kit-8.

Article Snippet: After-ward, 10 µL/well CCK-8 solutions were added, incubated for 1 hour and absorbance was measured at 450 nm using DTX-880 multimode microplate reader (Beckman Coulter, Inc., Brea, CA, USA).

Techniques: Incubation, Inverted Microscopy, CCK-8 Assay, Cell Counting

Journal: International Journal of Nanomedicine

Article Title: Immunotoxicity of titanium dioxide nanoparticles via simultaneous induction of apoptosis and multiple toll-like receptors signaling through ROS-dependent SAPK/JNK and p38 MAPK activation

doi: 10.2147/IJN.S176087

Figure Lengend Snippet: Effects of TiO 2 NPs on the total ROS and NO production. Notes: ( A ) Total ROS generation. ( B ) NO production. RAW 264.7 cells were incubated with indicated concentrations of TiO 2 SA20(−) NPs for 24 hours. Then cells were washed twice and incubated with H 2 DCF-DA (10 µmol/L) for 30 minutes. Cells were washed and analyzed using a multimode microplate reader DTX-880. Untreated cells were considered as control in the experiment. Then, intracellular NO production was measured by Griess reagent system (Promega Corp.) using ELx800 microplate reader (BioTek, Thermo Scientific) at 540 nm. Data are presented as the mean ± standard error of mean; * P <0.05, ** P <0.01, and *** ,### P <0.001 indicate significant differences when tested with ANOVA. Tukey’s test was used for post hoc tests. Abbreviations: TiO 2 , titanium dioxide; NO, nitric oxide; NPs, nanoparticles; ROS, reactive oxygen species.

Article Snippet: After-ward, 10 µL/well CCK-8 solutions were added, incubated for 1 hour and absorbance was measured at 450 nm using DTX-880 multimode microplate reader (Beckman Coulter, Inc., Brea, CA, USA).

Techniques: Incubation