Journal: Nutrition & Metabolism

Article Title: Resveratrol attenuates endothelial oxidative injury by inducing autophagy via the activation of transcription factor EB

doi: 10.1186/s12986-019-0371-6

Figure Lengend Snippet: Sequences of primers used in quantitative RT-PCR

Article Snippet: Antibodies of P62, TEFB, and LAMP1 were obtained from Cell Signaling Technology, whereas, β-actin antibody, fluorescein isothiocyanate (FITC)-conjugated secondary antibody, TFEB siRNA and control siRNA were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Sequencing

Journal: Nutrition & Metabolism

Article Title: Resveratrol attenuates endothelial oxidative injury by inducing autophagy via the activation of transcription factor EB

doi: 10.1186/s12986-019-0371-6

Figure Lengend Snippet: RSV increases TFEB activity in HUVECs. HUVECs were treated with different concentrations (0.1, 1, 10 μM) of RSV for 2 h. a The levels of nuclear and cytoplasmic TFEB were analyzed by western blotting, Histone H3 and β-actin were the standards for protein loading, respectively. b The bar graph shows the protein level quantification of nuclear TFEB (n-TFEB), cytoplasmic TFEB (c-TFEB) and the ratio of the endogenous nuclear TFEB to cytoplasmic TFEB. c The mRNA level of TFEB was then analyzed by qRT-PCR. d The mRNA levels of TFEB target genes were measured using qRT-PCR. Values are presented as means ± SD ( n = 3); * p < 0.05, ** p < 0.01 compared with control groups

Article Snippet: Antibodies of P62, TEFB, and LAMP1 were obtained from Cell Signaling Technology, whereas, β-actin antibody, fluorescein isothiocyanate (FITC)-conjugated secondary antibody, TFEB siRNA and control siRNA were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Activity Assay, Western Blot, Quantitative RT-PCR, Control

Journal: Nutrition & Metabolism

Article Title: Resveratrol attenuates endothelial oxidative injury by inducing autophagy via the activation of transcription factor EB

doi: 10.1186/s12986-019-0371-6

Figure Lengend Snippet: RSV stimulated autophagy through activating TFEB in PA-treated HUVECs. Cells were pretreated with RSV (10 μM) for 2 h, then treatment with PA (200 μM) for another 24 h. Furthermore, TFEB was knocked-down by siRNA transfection in HUVECs as described in the Materials and methods section. a The mRNA levels of TFEB was detected in HUVECs by using qRT-PCR. b - c The bar graph shows the protein level quantification of nuclear TFEB (n-TFEB), cytoplasmic TFEB (c-TFEB) and the ratio of the endogenous nuclear TFEB to cytoplasmic TFEB were analyzed by western blotting ( b ) and the ratio displayed by bar graph ( c ). d - e The protein levels of LC3 I, LC3 II, P62 and LAMP1 were analyzed by western blotting ( d ) and quantification displayed by bar graph ( e ). f The mRNA levels of TFEB target genes were measured using qRT-PCR. g The cells were incubated with lysotracker red (50 nM, 15 min, at 37 °C) and immediately visualized by confocal microscopy. h The average LTR fluorescence was expressed as the MFI using IPP 6.0 software. Values are presented as means ± SD ( n = 3); * p < 0.05, ** p < 0.01 compared between the marked groups

Article Snippet: Antibodies of P62, TEFB, and LAMP1 were obtained from Cell Signaling Technology, whereas, β-actin antibody, fluorescein isothiocyanate (FITC)-conjugated secondary antibody, TFEB siRNA and control siRNA were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Transfection, Quantitative RT-PCR, Western Blot, Incubation, Confocal Microscopy, Fluorescence, Software

Journal: Nutrition & Metabolism

Article Title: Resveratrol attenuates endothelial oxidative injury by inducing autophagy via the activation of transcription factor EB

doi: 10.1186/s12986-019-0371-6

Figure Lengend Snippet: RSV attenuated oxidative injury through stimulating autophagy in a TFEB-dependent manner in PA-treated HUVECs. Cells were pretreated with RSV (10 μM) for 2 h, then treatment with PA (200 μM) for another 24 h. Furthermore, TFEB was knocked-down by siRNA transfection in HUVECs as described in the Materials and methods section. a - c Intracellular ROS levels were estimated using a probe DCFH-DA by confocal microscopy ( a ) and bar charts show quantification of the ROS generation levels expressed as the fold change ( b ), and by microplate reader, fluorescence was read at 485 nm for excitation and 520 nm for emission ( c ). d The levels of MDA were detected using an MDA detection kit. e The cell viability was measured using a CCK-8 detection kit, Values are presented as means ± SD ( n = 6); * p < 0.05, ** p < 0.01 compared between the marked groups

Article Snippet: Antibodies of P62, TEFB, and LAMP1 were obtained from Cell Signaling Technology, whereas, β-actin antibody, fluorescein isothiocyanate (FITC)-conjugated secondary antibody, TFEB siRNA and control siRNA were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Transfection, Confocal Microscopy, Fluorescence, CCK-8 Assay

Journal: Nutrition & Metabolism

Article Title: Resveratrol attenuates endothelial oxidative injury by inducing autophagy via the activation of transcription factor EB

doi: 10.1186/s12986-019-0371-6

Figure Lengend Snippet: Schema of RSV-induced autophagy via activating TFEB in PA-stimulated HUVECs. Depending on the proposed signaling pathways: RSV activates TFEB and TFEB-target genes. Further, inducing the entire autophagy progress, concluding autophagosome as well as lysosome formation, ultimately, RSV attenuates PA-caused endothelial oxidative injury

Article Snippet: Antibodies of P62, TEFB, and LAMP1 were obtained from Cell Signaling Technology, whereas, β-actin antibody, fluorescein isothiocyanate (FITC)-conjugated secondary antibody, TFEB siRNA and control siRNA were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Protein-Protein interactions