Journal: World Journal of Gastroenterology

Article Title: Evaluation of a 3,5-diethoxycarbonyl-1,4-dihydrocollidine diet-induced mouse model in a comparative experimental study of portal hypertension

doi: 10.3748/wjg.v32.i9.114207

Figure Lengend Snippet: The 3,5-diethoxycarbonyl-1,4-dihydrocollidine diet induces abnormal proliferation of intrahepatic vascular structures and extrahepatic portosystemic shunting. A: Immunohistochemistry (IHC) staining for vascular endothelial growth factor receptor 2, cluster of differentiation 31, and vascular endothelial growth factor A in mouse livers; B-J: Statistical analysis of positive staining areas from IHC results; K-M: Hematoxylin-eosin staining of mesenteric tissues. Sample sizes ( n ): 8:8 (normal chow diet vs 3,5-diethoxycarbonyl-1,4-dihydrocollidine), 8:7 (sham vs bile duct ligation), and 8:7:8 (oil vs 8-week carbon tetrachloride vs 12-week carbon tetrachloride). b P < 0.01. c P < 0.001. NS: No significant; NCD: Normal chow diet; DDC: 3,5-diethoxycarbonyl-1,4-dihydrocollidine; BDL: Bile duct ligation; CCl 4 : Carbon tetrachloride; CD: Cluster of differentiation; VEGFR2: Vascular endothelial growth factor receptor 2; VEGF-A: Vascular endothelial growth factor A.

Article Snippet: Primary antibodies against collagen 1 (1:2500, catalog No. 67288-1-Ig, Proteintech), α-smooth muscle actin (SMA) (1:200, catalog No. ab5694, Abcam), desmin (1:4000, catalog No. 16520-1-AP, Proteintech), lymphatic vessel endothelial hyaluronan receptor 1 (LyVE-1) (1:100, catalog No. ab219556, Abcam), cluster of differentiation (CD) 34 (1:1000, catalog No. 14486-1-AP, Proteintech), von Willebrand factor (vWF) (1:200, catalog No. 27186-1-AP, Proteintech), vascular endothelial growth factor receptor 2 (VEGFR2) (1:150, catalog No. ab2349, Abcam), vascular endothelial growth factor A (VEGF-A) (1:100, catalog No. ab52917, Abcam), and CD31 (1:5000, catalog No. 11265-1-AP, Proteintech) were applied overnight at 4 °C, followed by 60-minute incubation with secondary antibodies at room temperature.

Techniques: Immunohistochemistry, Staining, Ligation

Journal: Cellular and Molecular Immunology

Article Title: Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line

doi: 10.1038/s41423-020-0440-9

Figure Lengend Snippet: Isolation of VEGF-A-neutralizing mAbs. a The sorting experiment to enhance the affinities of the anti-hVEGF cells. The C018 cells cultured without TSA were fluorescently stained with anti-hIgG and hVEGF. The cells showing higher reactivity to the antigen than the major population (gate “P3”) were singly sorted (left). The reactivity of the clones before and after affinity maturation was analyzed by flow cytometry (blue, parental C018; red, C018AM-20, the representative clone after affinity maturation) (right). b Bar chart representing the effects of anti-VEGF-A mAbs on the proliferation of HUVECs against the Ab(−) control. VEGF-A and serially diluted anti-hVEGF-A mAbs were preincubated and added to the HUVEC culture. The number of cells was counted after two days. P values represent significant differences between the cells cultured in the presence of anti-VEGF mAbs and identical concentrations of the negative control (anti-TNFα) mAb; error bars represent ±s.d. ( n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001. c SPR analysis of the anti-hVEGF mAbs. Serially diluted hVEGF-A was applied to the anti-hVEGF-A mAb immobilized on biosensor chips. d Summary of antibody binding constants. Association rate (ka) and dissociation rate (kd) constants were determined by SPR analysis. Dissociation constant (KD) values were calculated as kd/ka. e The sequences of the anti-hVEGF-A V H regions containing VH3-23-derived sequences. The sequences of B021, B015, C018 and C018AM-20 are compared with the original synthetic VDJ sequence designed for SCLs. The colored boxes indicate the original V genes: V3-23 (green), D3-3 (purple), JH4b (brown). The red and blue horizontal bars represent GC tracts (corresponding pseudogenes are described beside the bars) and SHM, respectively

Article Snippet: Recombinant human VEGF R2/KDR Fc chimera protein (R&D Systems) was immobilized onto MaxiSorp 384-well plates at 62.5 ng/well at 4 °C overnight.

Techniques: Isolation, Cell Culture, Staining, Clone Assay, Flow Cytometry, Control, Negative Control, Binding Assay, Derivative Assay, Sequencing

Journal: Molecular and cellular biochemistry

Article Title: Sex-Dimorphic Moderate Hypoglycemia Preconditioning Effects on Hippocampal CA1 Neuron Bio-Energetic and Anti-Oxidant Function

doi: 10.1007/s11010-020-03806-7

Figure Lengend Snippet: Summary of Moderate Hypoglycemia Preconditioning (MH) Effects on Hippocampal CA1 Neuron Glycolytic, Tricarboxylic Acid, Oxidative Respiratory, and Anti-Oxidative Stress Enzyme Responses to Severe Hypoglycemia (SH) in Male versus Female Rats

Article Snippet: Technol.), goat [pyruvate dehydrogenase kinase component E1 (PDK1; CA-46155, 1:1250; ProSci.), IDH2 (CA-42704, 1:1500; ProSci.), ATPs (CA-42704, 1:1500; ProSci.), 4-HNE (NB100–63093, 1:3000, Novus Biol.), or mouse [NT (MAB3248; 1:2000; R&D Research, MN)], Membranes were incubated for 1 hr with peroxidase-conjugated rabbit anti-goat (AP106P, 1:5000; EMD Millipore, Billerica, MA), goat anti-rabbit (NEF812001EA, 1:5000; PerkinElmer, Boston, MA), or goat anti-mouse (NEF822001EA; 1:5000; PerkinElmer) secondary antibodies before exposure to SuperSignal West Femto chemiluminescent substrate (34095; ThermoFisherScientific, Rockford, IL).

Techniques:

Journal: Molecular and cellular biochemistry

Article Title: Sex-Dimorphic Moderate Hypoglycemia Preconditioning Effects on Hippocampal CA1 Neuron Bio-Energetic and Anti-Oxidant Function

doi: 10.1007/s11010-020-03806-7

Figure Lengend Snippet: For each treatment group, triplicate CA1 lysate pools were created for Western blot analysis of AMPK (Figure 5A: male data at left, female data at right), pAMPK (Figure 5B), 4-HNE (Figure 5D), or NT (Figure 5E). Figure 5C presents mean pAMPK/AMPK ratios in male versus female CA1 neurons. Data illustrate mean normalized protein O.D. measures ± S.E.M. for male and female V1−4 (solid white bars), V1−3/SH4 (solid gray bars), and MH1−3/SH4 (cross-hatched gray bars) treatment groups. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: Technol.), goat [pyruvate dehydrogenase kinase component E1 (PDK1; CA-46155, 1:1250; ProSci.), IDH2 (CA-42704, 1:1500; ProSci.), ATPs (CA-42704, 1:1500; ProSci.), 4-HNE (NB100–63093, 1:3000, Novus Biol.), or mouse [NT (MAB3248; 1:2000; R&D Research, MN)], Membranes were incubated for 1 hr with peroxidase-conjugated rabbit anti-goat (AP106P, 1:5000; EMD Millipore, Billerica, MA), goat anti-rabbit (NEF812001EA, 1:5000; PerkinElmer, Boston, MA), or goat anti-mouse (NEF822001EA; 1:5000; PerkinElmer) secondary antibodies before exposure to SuperSignal West Femto chemiluminescent substrate (34095; ThermoFisherScientific, Rockford, IL).

Techniques: Western Blot