Journal: Iranian Journal of Basic Medical Sciences

Article Title: Aptamer-functionalized mesenchymal stem cells-derived exosomes for targeted delivery of SN38 to colon cancer cells

doi: 10.22038/IJBMS.2023.68039.14873

Figure Lengend Snippet: (A) Scanning electron microscope (SEM) image of exosomes with scale bar 200 nm, (B) Scanning electron microscope (AFM) images of exosomes, (C) Size of exosomes with dynamic light scattering (DLS)

Article Snippet: Thereafter, the droplet was thoroughly dried at 25°C for 12 hr to collect the images by the employment of an atomic force microscope (Nano Wizard II Nanoscience AFM, JPK Instruments, Germany) ( , ).

Techniques: Microscopy

Journal: Antioxidants (Basel, Switzerland)

Article Title: Activation of NRF2 Signaling Pathway Delays the Progression of Hyperuricemic Nephropathy by Reducing Oxidative Stress.

doi: 10.3390/antiox12051022

Figure Lengend Snippet: Figure 10. Mechanisms by which the activation of NRF2 signaling ameliorates kidney injury in HN mice. Under physiological conditions, cells produce an appropriate amount of ROS to adapt to normal physiological activities, while NRF2 exists in the cytoplasm at a lower level. High levels of UA will be transported into cells by transporters on the apical membrane of renal tubular epithelial cells, stimulating NOX4 to produce excessive ROS. Excessive ROS causes mitochondrial fusion and fission disorders, leading to mitochondrial dysfunction. Damaged mitochondria produce excessive mitochondrial ROS and aggravate cellular oxidative stress. The administration of NRF2 agonist sulforaphane (SFN) can improve cellular oxidative stress by down-regulating NOX4 expression and remodeling mitochondrial homeostasis. Moreover, NRF2 activation enhances the antioxidant capacity of cells by upregulating the expression of HO-1/NQO1, reduces renal fibrosis by downregulating the expression of TGF-β1/α-SMA/Collagen 1, and ultimately improves renal function and slows down disease progression.

Article Snippet: The membranes were blocked with blocking buffer (TBS, 0.1% Tween-20, 5% non-fat milk or 2% BSA) for 2 h at room temperature and incubated with Antioxidants 2023, 12, 1022 6 of 25 primary antibodies against NOX4 (ABclonal, Wuhan, China, Cat# A11274, 1:1000 dilution), HO-1 (Proteintech, Wuhan, China, Cat# 10701-1-AP, 1:1000 dilution), NQO1 (Proteintech, Wuhan, China, Cat# 67240-1-Ig, 1:1000 dilution), MFN1 (ABclonal, Wuhan, China, Cat# A9880, 1:1000 dilution), MFN2 (ABclonal, Wuhan, China, Cat# A19678, 1:1000 dilution), FIS1 (ABclonal, Wuhan, China, Cat# A19666 1:1000 dilution), TGF-β1 (Proteintech, Wuhan, China, Cat# 21898-1-AP, 1:1000 dilution), α-SMA (Proteintech, Wuhan, China, Cat# 14395-1-AP, 1:5000 dilution), collagen 1 (Proteintech, Wuhan, China, Cat# 67288-1-Ig, 1:5000 dilution), and GAPDH (ABclonal, Wuhan, China, Cat# AC001, 1:10,000 dilution) at 4 ◦C overnight, respectively.

Techniques: Activation Assay, Membrane, Expressing, Biomarker Discovery

Journal:

Article Title: NMR Mapping and Functional Confirmation of the Collagen Binding Sites of MMP-2 †

doi: 10.1021/bi900513h

Figure Lengend Snippet: Panel A. HSQC spectra of 0.5 mM 15N-labeled CBD in 25 mM sodium phosphate buffer, 5% D2O/H2O, pH 7.0 were recorded before (black) and after titration with collagen peptide P713 at concentrations of 0.15 mM (red), 0.3 mM (green), and 1.5 mM (blue). Residues that exhibited shift perturbations of backbone amides upon P713 peptide addition are shown and labeled with their one letter amino acid code and residue number. Residues which were selected for alanine substitution are underlined. Residues selected as negative controls are labeled in italics. The insert shows the change in the chemical shift of F297 as P713 is added. Panel B. The concentration dependent chemical shifts of three representative CBD residues upon binding P713 were plotted against the concentration of P713. Panel C. Magnitudes of chemical shift change (Δδ) for CBD residues in the presence of a P713 concentration of 1.5 mM. Chemical shift perturbations, Δδ, were calculated from proton and nitrogen shift by using the formula {[(δΔH)2+(δΔN/5)2]/2}½. Panel D. Three-dimensional surface representation of individual CBD modules showing the positions of residues that underwent shift changes in the presence of P713. The intensity of blue color in each CBD modules is proportional to the magnitude of the chemical shifts changes induced by peptide P713. The residues are labeled with the one letter amino acid code and residue number. The structure shown is from the reported MMP-2 crystal structure, PDB 1CK7.

Article Snippet: The 12 residue-long collagen peptide P713 ( 25 ) (CGA(HYP)GA(HYP)GSQGA) was synthesized at the UTHSCSA Protein Core Facility by sequential addition of FMOC protected amino acids on a Multiple Peptide Synthesizer Model 396 MPS (Advanced Chemtech, Louisville, KY).

Techniques: Labeling, Titration, Concentration Assay, Binding Assay

Journal:

Article Title: NMR Mapping and Functional Confirmation of the Collagen Binding Sites of MMP-2 †

doi: 10.1021/bi900513h

Figure Lengend Snippet: Panel A. CBD variants were purified from E. coli and dissolved at concentrations of 150–200 µM in 25 mM phosphate buffer, pH 7.0 with 5% D2O. One-dimensional 1H NMR spectra were collected at a frequency of 700 MHz. Typically, signal dispersion above 8.5 ppm representing backbone amide groups and below 0.8 ppm representing methyl groups occurs only if the protein is structurally ordered. Alanine substitution of key residues of rCBD variants showed that their overall fold was not significantly perturbed compared with wildtype rCBD. Four representative variant are presented. Panel B. Binding of wildtype (WT) and CBD variants to peptide P713 by surface plasmon resonance. Alkylated CBD (AlkCBD) which did not bind gelatin served as negative control. P713 was immobilized on SPR CM5 chips, and 1 µM purified WT and variant CBDs in 10 mM HEPES, 150 mM NaCl, 3 mM EDTA were then passed over the peptide surfaces. Bar graph presents mean and SD (bars) of specific binding determined from Biacore sensorgrams and expressed in response units (RU). Alk-CBD represents non-functional alkylated WT CBD.

Article Snippet: The 12 residue-long collagen peptide P713 ( 25 ) (CGA(HYP)GA(HYP)GSQGA) was synthesized at the UTHSCSA Protein Core Facility by sequential addition of FMOC protected amino acids on a Multiple Peptide Synthesizer Model 396 MPS (Advanced Chemtech, Louisville, KY).

Techniques: Purification, Variant Assay, Binding Assay, SPR Assay, Negative Control, Functional Assay