Incubation:
Article Title: DNA methylation-mediated alterations in Copper(I/II) redox equilibrium underlie lead-induced neurotoxicity.
Article Snippet: Lead (Pb), a ubiquitous environmental toxin, poses significant risks to central nervous system health, primarily by disrupting essential metal homeostasis in the brain.. While epigenetic regulation and proteomic expression are significantly affected by Pb, its specific molecular impact on copper (Cu) redox states remains poorly understood.. This study systematically investigated the molecular mechanisms underlying Pb-induced neurotoxicity in SHSY5Y cells through integrated epigenomics and proteomics analysis.
Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Article Title: Self-illuminating activatable chemotherapy enabled by pH-MPO-ROS cascade-amplified nanoparticles for cancer and metastatic lymph nodes
Article Snippet: Lymph nodes (LNs) are the primary site of metastasis for various cancers spreading through the lymphatic system.. The role of lymph node dissection, as outlined by clinical guidelines, has increasingly been called into question.. Here, we developed a self-illuminating and pH-MPO-ROS cascade-activatable paclitaxel nanoparticle (PCAPN), which integrates ultra-pH-sensitive, self-illuminated imaging, in situ cascade gated activation of therapeutic agents to specifically eliminate the residual tumors and metastatic LNs after surgery.
Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation.
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Cell Culture:Article Title: A multifunctional exudate-absorptive patch accelerates burn wound healing via antioxidant, anti-inflammatory and immunomodulatory effects
Article Snippet: The surface chemical bonds of patches were analyzed by Fourier transform infrared spectroscopy (FTIR, Spectrum3, Perkinelmer, USA) with a scanning range of 400-4000 cm-1. .. 2.3 Scavenging effect of SPC@C on intracellular ROS RAW264.7 cells were seeded in confocal dishes (NEST Biotechnology, cat. no. 801002) at a density of 3 × 105 cells/mL and cultured for 24 h. The culture medium was replaced with complete DMEM containing lipopolysaccharide (LPS, 100 ng/mL, Servicebio, China) and SPC@C (diameter: 5 mm, thickness: 2 mm), with the materials placed above the cells (without direct contact). ..
Transfection:
Blocking Assay:Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation.
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Staining:Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation.
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Labeling:Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation.
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Fluorescence:Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation.
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Microscopy:Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
Article Title: Activating NEDD4L suppresses EGFR-driven lung adenocarcinoma growth via facilitating EGFR proteasomal degradation.
Article Snippet: .. Cells were plated in confocal dishes (NEST Biotechnology) and incubated to 50% confluency, then washed with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with QuickBlockTM Blocking Buffer for Immunol Staining (Beyotime, Shanghai, China) for 1 h at room temperature, and incubated with desired primary antibodies against EGFR (Cat. #4267, Cell Signaling Technology; 1:100), NEDD4L (Cat. #13690–1-AP, Proteintech; 1:200), HA-tag (Cat. #2367/3724, Cell Signaling Technology; 1:100) at 4 °C overnight, followed by incubation of fluorochrome-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature under dark, finally labeled by using DAPI (Beyotime, Shanghai, China) for 5 min, and subjected to a confocal fluorescence microscope (ZEISS,Jena,Germa). ..
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