Journal: Molecules

Article Title: Antioxidant and Anti-Inflammatory Activities of a Natural Compound, Shizukahenriol, through Nrf2 Activation

doi: 10.3390/molecules200915989

Figure Lengend Snippet: Figure 3. Induction of antioxidant enzyme gene expression by SZH in BV-2 microglial cells (a) effect of SZH on BV-2 microglial cell viability using CCK-8 cytotoxicity assay for 24 h; (b) expression kinetics of HO-1, a major nrf2-inducing enzyme, by SZH treatment; (c–e) BV-2 microglial cells were harvested after 24 h, the cell lysate were subjected to Western blot; (c) HO-1 (d) GCLM; and (e) GCLC, densitometric analyses were performed and the data were normalized against the internal control β-actin. These data are expressed as fold induction of untreated control ± S.E.M. * p < 0.05, ** p < 0.01 or *** p < 0.005 vs. DMSO only treated control.

Article Snippet: The membranes were incubated overnight with primary antibody against NRF2 (D1Z9C; Cell signaling, Denvers, MA, USA, 1:300), HO-1 (Enzo Life Sciences, Ann Arbor, MI, USA, 1:1000), GCLC (Novus Biologicals, Littleton, CO, USA, 1:1000), GCLM (FL-274; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA, 1:1000), iNOS (EPR16635; Abcam, Cambridge, UK, 1:500), NF-κB p65 (C22B4; Cell signaling, Denvers, MA, USA, 1:1000), LAMIN-B1 (D4Q4Z; Cell Signaling, 1:1000), and β-ACTIN (N-21; Santa Cruz Biotechnology, Inc., 1:1000) at 4 °C.

Techniques: Gene Expression, CCK-8 Assay, Cytotoxicity Assay, Expressing, Western Blot, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: New generation of DNA-based immunotherapy induces a potent immune response and increases the survival in different tumor models

doi: 10.1136/jitc-2020-001243

Figure Lengend Snippet: B16F10-OVA therapeutic vaccination comparing pTOP vaccines and classical DNA vaccination, and combination with immune checkpoint blockade. (A) Schematic protocol of the B16F10-OVA injection and therapeutic DNA vaccination. C57BL/6 mice were first injected with B16F10-OVA. The DNA vaccines were intramuscularly electroporated 2, 9 and 16 days after injection of the tumor cells. (B) Tumor growth curves for the different groups. (C) percentage of survival as a function of time. The error bars represent the mean±SEM; n=6. Statistical analysis: one-way ANOVA with Tukey’s multiple comparisons test, two-way ANOVA with Bonferroni post-tests or Mantel-Cox test for comparison of survival curves. *p<0.05, compared with naive or to the specified group. (D) schematic protocol of the B16F10-OVA injection, therapeutic DNA vaccination and immune checkpoint blockade administration. C57BL/6 mice were first injected with B16F10-OVA. the DNA vaccines were intramuscularly electroporated 2, 9 and 16 days after injection of the tumor cells. Immune checkpoint blockade antibodies against CTLA4 (100 µg) and PD1 (100 µg) were injected intraperitoneally 3, 6 and 9 days after tumor injection. (E) tumor growth curves for the different groups. (F) percentage of survival as a function of time. The error bars represent the mean±SEM; n=6. Statistical analysis: One-way ANOVA with Tukey’s multiple comparisons test, two-way ANOVA with Bonferroni post-tests or Mantel-Cox test for comparison of survival curves. **p<0.01, compared with naive or to the specified group. ANOVA, analysis of variance; MST, median survival time; OVA, ovalbumin; pOVA, plasmid OVA; pTOP, plasmid to deliver T cell epitopes; pVSVG, plasmid vesicular stomatitis virus glycoprotein.

Article Snippet: Immune checkpoint blockade antibodies directed against CTLA4 (clone 9D9) and PD1 (clone 29 F.A12) were purchased from Bioconnect (Netherlands) and mice were injected intraperitoneally with 100 μg of each antibody in 100 μl of PBS 3, 6 and 9 days after tumor injection.

Techniques: Vaccines, Injection, Comparison, Plasmid Preparation, Virus