Journal: Veterinary Sciences

Article Title: Roles of WNT6 in Sheep Endometrial Epithelial Cell Cycle Progression and Uterine Glands Organogenesis

doi: 10.3390/vetsci8120316

Figure Lengend Snippet: Details of antibodies.

Article Snippet: β-catenin , 51067-2-AP , Proteintech, Wuhan, China , _ , 1:5000 , β-Catenin, also known as CTNNB1, is a key downstream component of the canonical Wnt pathway that plays diverse and critical roles in embryonic development and adult tissue homeostasis..

Techniques: Quantitative Proteomics, Transformation Assay, Gene Expression

Journal: Veterinary Sciences

Article Title: Roles of WNT6 in Sheep Endometrial Epithelial Cell Cycle Progression and Uterine Glands Organogenesis

doi: 10.3390/vetsci8120316

Figure Lengend Snippet: The effects of WNT6 transfection on β-catenin expression in EECs. ( A , B ) Relative expression of WNT6 and β-catenin protein to β-actin by WNT6 knockdown. ( C , D ) Relative expression of WNT6 and β-catenin protein to β-actin by WNT6 overexpression. si-WNT6, the small interfering RNA for WNT6 . pEX-4-WNT6, pEX-4 vector with WNT6 -CDs inserted. siRNA with scrambled sequence absent in the sheep genome and pEX-4 was severed as negative control for knockdown and overexpression, respectively. The values are presented as means ± SDs. Means with different letters indicate significant differences ( p < 0.05).

Article Snippet: β-catenin , 51067-2-AP , Proteintech, Wuhan, China , _ , 1:5000 , β-Catenin, also known as CTNNB1, is a key downstream component of the canonical Wnt pathway that plays diverse and critical roles in embryonic development and adult tissue homeostasis..

Techniques: Transfection, Expressing, Knockdown, Over Expression, Small Interfering RNA, Plasmid Preparation, Sequencing, Negative Control

Journal: iScience

Article Title: Bombinin-BO1 induces hepatocellular carcinoma cell-cycle arrest and apoptosis via the HSP90A-Cdc37-CDK1 axis

doi: 10.1016/j.isci.2024.110382

Figure Lengend Snippet:

Article Snippet: Materials purchased from Proteintech Group (Wuhan, China) included antibodies against GAPDH (proteintech, 10494-1-AP, 1:5000), Beta Actin (proteintech, 66009-1-Ig, 1:1000), goat anti-rabbit IgG (proteintech, 66912-1-lg, 1:10000), Pro Caspase-8 (Abcam, ab108333, 1:10000), Caspase-9 (proteintech, 10380-1-AP, 1:1000), Cleaved Caspase-9 (Abcam, ab2324, 1 ug/mL), caspase-3 (proteintech, 19677-1-AP, 1:2000), cleaved caspase-3 (Abcam, ab2302, 1:100), Bcl-2 (proteintech, 12789-1-AP, 1:1000), Bax (proteintech, 50599-2-lg,1:1000), HSP90A (proteintech, 60318-1-Ig, 1:5000), Cdc37 (proteintech, 10218-1-AP, 1:200), CDK1 (proteintech, 19532-1-Ig, 1:200), Cyclin A2 (proteintech, 18202-1-AP, 1:5000).

Techniques: Recombinant, Magnetic Beads, RNA Extraction, Quantitation Assay, Protein Concentration, Software, Lysis

Journal: eLife

Article Title: The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms

doi: 10.7554/elife.97017

Figure Lengend Snippet: Figure 4. The distribution and organization of NiV-F constructs in VLPs. (A) The incorporation of F-WT and mutants in VLPs. NiV-M-GFP, G-HA, and FLAG-tagged F-WT or mutants were transfected to 293T cells. The supernatants were collected at 48 hr post-transfection and analyzed on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) followed by western blotting. NiV-M was probed by polyclonal goat anti-GFP, NiV-G polyclonal rabbit anti-HA, F0 and F2 M2 monoclonal mouse anti-FLAG antibody. (B) Cross-section (Δz = 100 nm) of single-molecule localization microscopy (SMLM) images of the FLAG-tagged NiV-F-WT (WT), L53D, V108D, and Q393L on individual VLPs. Scale bar: 0.2 μm. (C) The classification of the ordered sequence of reachability distances of the NiV-F constructs localizations. Orange: F-WT (n = 306) and Q393L (n = 323); blue: L53D (n = 310) and V108D (n = 329). n is the number of VLPs used for classification analysis.

Article Snippet: DOI: https://doi.org/10.7554/eLife.97017 16 of 23 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Chemical compound, drug HEPES Fisher Scientific 15- 630- 080 25 mM for fusion buffer Chemical compound, drug Glutamine Fisher Scientific 25- 030- 081 2 mM for fusion buffer Chemical compound, drug CaCl2 Sigma- Aldrich C7902 1 mM for fusion buffer Chemical compound, drug Probenecid Sigma- Aldrich P8761- 25G 2.5 mM for fusion buffer Chemical compound, drug Solution D Fisher Scientific K1156 1:20 for Entry kinetics loading solution Antibody Anti- FLAG mouse monoclonal Sigma- Aldrich F1804 IF, SMLM: 1:100, Flow: 1:200; WB: 1:500–1:5000 Antibody Anti- HA rabbit polyclonal antibody Biolegend 902301 IF: 1:900 WB: 1:2000 Antibody Anti- GFP goat polyclonal antibody Abcam Ab5450 WB: 1:1000 Antibody Anti- mCherry rabbit polyclonal antibody Abcam Ab167453 WB: 1:2000 Antibody Anti-β-lactamase mouse monoclonal Santa Cruz Biotechnology Sc- 66062 WB: 1:1000 Antibody Anti- mouse donkey polyclonal antibody, Alexa Fluor 647 conjugated Invitrogen A31571 IF and SMLM: 1:400 Antibody Anti- rabbit donkey polyclonal antibody, Alexa Fluor 488 conjugated Invitrogen A21206 IF: 1:400 Antibody Anti- mouse donkey polyclonal antibody, Alexa Fluor 488 conjugated Invitrogen A21202 Flow: 1:400 Antibody Anti- goat donkey polyclonal antibody, HRP conjugated Jackson Immunoresearch 705- 035- 147 WB: 1:5000 Antibody Anti- mouse goat polyclonal antibody, HRP conjugated Bio- Rad 1705047 WB: 1:5000 Antibody Anti- rabbit goat polyclonal antibody, HRP conjugated Bio- Rad 1706515 WB: 1:5000 Commercial assay or kit μMACS anti- DYKDDDDK starting kit Miltenyi Biotec 130- 101- 636 Commercial assay or kit Renilla Luciferase Assay System Promega E2820 Commercial assay or kit LiveBLAzer FRET- B/G Loading Kit with CCF2- AM Invitrogen K1032 Commercial assay or kit QIAamp Viral RNA Kits for RNA Extraction QIAGEN 52904 Commercial assay or kit SuperScript III First- Strand Synthesis System Invitrogen 18080051 Software SMLM image reconstruction Liu et al., 2018 MATLAB Codes are available upon request.

Techniques: Construct, Transfection, Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot, Microscopy, Sequencing

Journal: Neuron

Article Title: Multimodal Single-Cell Analysis Reveals Physiological Maturation in the Developing Human Neocortex

doi: 10.1016/j.neuron.2019.01.027

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Biocytin was detected by Streptavidin Dylight 549 (1:500, Vector Labs SA-5549).

Techniques: Plasmid Preparation, RNAscope 2.5 HD Assay, RNA Sequencing Assay, Software

Journal: Science Progress

Article Title: Role of thrombus-derived exosomal lncRNA LOC101928697 in regulating endothelial function via FUS protein interaction in myocardial infarction

doi: 10.1177/00368504251372111

Figure Lengend Snippet: Analysis of the correlation between FUS protein and myocardial infarction. (a) Enrichment Analysis Bar Plot based on differential gene expression profiles in lncRNA microarray analysis.(b) Detection information about lncRNA LOC101928697 binding to FUS proteins in AnnoLnc2 database. (c) Detection information about lncRNA LOC101928697 binding to FUS protein in RBPDP database. (d) Scores in the RPISeq database on the model of lncRNA LOC101928697 binding to FUS protein. (e-g) Prediction information about lncRNA LOC101928697 binding to FUS protein in catRAPID website, (e) Statistical map information about protein and RNA binding sites, (f) Total scoring information, and (g) Interaction map showing the interaction region between protein and RNA. (h-i) Analyses about bioinformatics techniques based on GSE163772 in the GEO database, where (h) is a statistical map of FUS gene expression in endothelial cells of a mouse model of myocardial infarction, and (i) A scatter plot about the correlation between the level of FUS gene expression and the disease state (control vs. myocardial infarction).

Article Snippet: After extensive washing, the bound proteins were eluted, separated by SDS-PAGE, and analyzed by Western blot using anti-FUS antibody (Proteintech, Cat No. 11570-1-AP, dilution 1:5000) to detect the enrichment of FUS protein.

Techniques: Gene Expression, Microarray, Binding Assay, RNA Binding Assay, Control

Journal: Science Progress

Article Title: Role of thrombus-derived exosomal lncRNA LOC101928697 in regulating endothelial function via FUS protein interaction in myocardial infarction

doi: 10.1177/00368504251372111

Figure Lengend Snippet: Interaction of exosomal lncRNA LOC101928697 with FUS proteins. (a and b) The western blot detection of FUS protein expression in each group of cells and the statistical graph. (c) Statistical graph of RT-qPCR to detect the expression of FUS at the mRNA level in each group of cells. (d) The fluorescence graph of fluorescence in situ hybridization (FISH) experiment. In which FUS was labeled with green fluorescence, lncRNA LOC101928697 was labeled with red fluorescence, and the nucleus was labeled with blue fluorescence (20×). (e) Western blot detection of FUS protein following RNA pull-down using sense or antisense LOC101928697 transcripts. (f) Quantification of FUS protein enrichment in sense RNA pull-down versus antisense control, based on densitometric analysis. (g-h) Western blot detection of FUS protein expression in each group of cells after knockdown or overexpression of lncRNA LOC101928697 and the statistical graphs. (i) Statistical graph of mRNA level expression of FUS in each group of cells after knockdown or overexpression of lncRNA LOC101928697 by RT-qPCR assay. a p < 0.05 compared to control group. b p < 0.05 compared to exosome group. c p < 0.05 compared to siRNA + exosome group.

Article Snippet: After extensive washing, the bound proteins were eluted, separated by SDS-PAGE, and analyzed by Western blot using anti-FUS antibody (Proteintech, Cat No. 11570-1-AP, dilution 1:5000) to detect the enrichment of FUS protein.

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Fluorescence, In Situ Hybridization, Labeling, Protein Enrichment, Control, Knockdown, Over Expression