Journal: Cell

Article Title: In Vivo Target Gene Activation via CRISPR/Cas9-Mediated Trans -Epigenetic Modulation

doi: 10.1016/j.cell.2017.10.025

Figure Lengend Snippet: (A) Schematic of AAVs administration to Cas9 mice via tail-vein injection to prevent cisplatin-induced acute kidney injury (AKI). (B) qRT-PCR analyses of Il10 and klotho expression in liver tissues from mice injected with AAV-dgIL-10-MPH (n = 3) or AAV-dgKlotho-MPH (n = 3). Fold enrichments were calculated relative to dgMock controls. (C) AAV-dgKlotho-MPH increased serum levels of Klotho protein relative to dgMock controls in cisplatin-treated mice (n = 6). Data are means ± SD. (D) Blood urea nitrogen (BUN) and serum creatinine (S-cre) levels in cisplatin-induced AKI mice were reduced by AAV-mediated IL-10 or Klotho overexpression. (E and F) Histological sections from indicated mice were subjected to H&E and PAS staining (E) and pathological features were quantified (F) (10~15 slides analyzed for 3–4 mice per group). Scale bar, 50 µm. (G) Survival curves of mice treated with the indicated AAV-dgRNAs for 8 days followed by a 17 g/Kg cisplatin challenge (via intraperitoneal injection; n = 8 for each group). See also Figure S3.

Article Snippet: Mouse sera was subjected to ELISA assay following the standard protocol (Mouse Klotho ELISA kit, CUSABIO; Mouse IL-10 ELISA kit, Affymetrix eBioscience; Mouse Insulin ELISA kit, ALPCO).

Techniques: Injection, Quantitative RT-PCR, Expressing, Over Expression, Staining

Journal: Scientific Reports

Article Title: Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats

doi: 10.1038/s41598-017-11260-7

Figure Lengend Snippet: Comparison of systemic factors using enzyme-linked immunosorbent assays (ELISA). Brain-derived nerve growth factor (BDNF) was upregulated after both AE and NMES, with the most dramatic change evident after a medium dose of NMES resulting in an almost 8-fold increase. Little change was evident in VEGF-A, with only the low AE condition showing a minor non-significant 6% increase. The low AE condition also increased IGF-1 by 33% and showed an AE dose-dependent decrease, with the medium condition being equivalent to control levels. NMES did not affect IGF-1 levels. In contrast, Klotho revealed a significant AE dose-dependent modulation of levels with a medium dose of AE doubling the amount of Klotho in the blood compared to controls. NMES also produced an increase of Klotho for the low and high condition, but not for the medium dose. (*p < 0.05; **p < 0.01).

Article Snippet: Brain-derived neurotrophic factor (BDNF, ERBDNF, Thermo scientific), Vascular endothelial growth factor-A (VEGF-A, RRV00, R&D), Insulin-like growth factor 1 (IGF-1, CSB-E04582r, CUSABIO) and Klotho (CSB-E14958r, CUSABIO) levels were determined by quantitative enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Derivative Assay, Control, Produced

Journal: Scientific Reports

Article Title: Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats

doi: 10.1038/s41598-017-11260-7

Figure Lengend Snippet: Correlations between biological variables and MCT.

Article Snippet: Brain-derived neurotrophic factor (BDNF, ERBDNF, Thermo scientific), Vascular endothelial growth factor-A (VEGF-A, RRV00, R&D), Insulin-like growth factor 1 (IGF-1, CSB-E04582r, CUSABIO) and Klotho (CSB-E14958r, CUSABIO) levels were determined by quantitative enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats

doi: 10.1038/s41598-017-11260-7

Figure Lengend Snippet: Contour maps – systemic factors. Based on the factorial design that arrayed treatment interventions to a control (0 dose), low, medium and high dose, contour plots of the dependent variables can be drawn to illustrate how these variables interact across the experimental space. It is evident here that systemic BDNF levels were most dramatically increased using NMES, but due to the lack of increase in MCT performance there was little interaction between treatment dose and % change in speed. BDNF did also not reveal a dramatic dose-dependent interaction for AE. A similar lack of interaction was evident for VEGF-A, with some evidence of an increase in levels for a low AE condition that results in a 25–50% change in speed. A clearer interaction between dosing and MCT was evident for AE on IGF-1. A low to medium dose of therapy here produced higher levels of factor release, but this was associated with a poorer performance. Lower levels of IGF-1 were actually induced by the high AE condition, producing the most significant performance change (upper right corner). Minimal interaction was seen for NMES, considering that there was less of an effect on MCT, demonstrating that IGF-1 levels in this paradigm were almost homogenous throughout experimental space. In the AE condition, the contour plot for Klotho indicated that the medium dose produced the highest level of Klotho release, which was associated with the highest level of behavioral change. In contrast the plot for NMES did not reveal a clear interaction between these variables.

Article Snippet: Brain-derived neurotrophic factor (BDNF, ERBDNF, Thermo scientific), Vascular endothelial growth factor-A (VEGF-A, RRV00, R&D), Insulin-like growth factor 1 (IGF-1, CSB-E04582r, CUSABIO) and Klotho (CSB-E14958r, CUSABIO) levels were determined by quantitative enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Control, Produced

Journal: Scientific Reports

Article Title: Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats

doi: 10.1038/s41598-017-11260-7

Figure Lengend Snippet: Hippocampal gene expression. BDNF mRNA was upregulated after AE, but did not exhibit a dose-dependency. A significant 10% increase in hippocampal BDNF was evident after low NMES and showed a dose dependent decrease to the level of controls. VEGF-A was upregulated in all NMES conditions, but did not change after a medium and high dose of AE. A low dose actually decreased VEGF-A expression. IGF-1 was upregulated in all conditions by approx. 17%. Only the low AE condition produced a lower 5% upregulation. Klotho revealed an expression profile similar to IGF-1 (27% increase), but with a more marked increase in expression in the low AE condition (12%). (*p < 0.05; **p < 0.01).

Article Snippet: Brain-derived neurotrophic factor (BDNF, ERBDNF, Thermo scientific), Vascular endothelial growth factor-A (VEGF-A, RRV00, R&D), Insulin-like growth factor 1 (IGF-1, CSB-E04582r, CUSABIO) and Klotho (CSB-E14958r, CUSABIO) levels were determined by quantitative enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Gene Expression, Expressing, Produced

Journal: Scientific Reports

Article Title: Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats

doi: 10.1038/s41598-017-11260-7

Figure Lengend Snippet: Correlations ELISA and mRNA.

Article Snippet: Brain-derived neurotrophic factor (BDNF, ERBDNF, Thermo scientific), Vascular endothelial growth factor-A (VEGF-A, RRV00, R&D), Insulin-like growth factor 1 (IGF-1, CSB-E04582r, CUSABIO) and Klotho (CSB-E14958r, CUSABIO) levels were determined by quantitative enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats

doi: 10.1038/s41598-017-11260-7

Figure Lengend Snippet: Contour maps – hippocampal gene expression. By accounting for the different levels of dosing for AE and NMES, contour maps were generated to expose interactions between variables in the experimental space. It was evident here that BDNF expression in the hippocampus was highest for the high AE and low NMES conditions. As there was no significant improvement in MCT performance from NMES, this produced a rather flat banding of expression levels. The highest BDNF levels were evident between no change to a 25% performance decrease. For AE, the highest levels were achieved by a high dose that resulted in the highest level of performance increase. VEGF-A revealed the highest increase in the NMES group with a low to a medium dosing leading to peak gene expression. This was associated with improvements in MCT. AE yielded a dose-dependent increase in IGF-1 signaling that was linked to improved performance with a diagonal pattern across the experimental space. A similar effect was evident for Klotho with lower levels being associated with poorer performance and higher levels indicating better performance. A medium dosing of AE is sufficient to achieve peak performance and high levels of Klotho. It was also highly upregulated in the NMES condition with only very high decreases in performance showing a low level of Klotho.

Article Snippet: Brain-derived neurotrophic factor (BDNF, ERBDNF, Thermo scientific), Vascular endothelial growth factor-A (VEGF-A, RRV00, R&D), Insulin-like growth factor 1 (IGF-1, CSB-E04582r, CUSABIO) and Klotho (CSB-E14958r, CUSABIO) levels were determined by quantitative enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Gene Expression, Generated, Expressing, Produced