Journal: Molecular Therapy. Nucleic Acids

Article Title: Inhibition of adenovirus replication by CRISPR-Cas9-mediated targeting of the viral E1A gene

doi: 10.1016/j.omtn.2023.02.033

Figure Lengend Snippet: Structure of the recombinant adenovirus vector constructs and gRNA sequences incorporated into them (A) Schematic of HAdV-5-based vectors with inserted individual gRNA sequences. All adenoviral vectors are based on the HAdV-5-derived vector pAd/PL-DEST (Thermo Fisher Scientific) and lack the E1 and E3 regions. Cas9 and gRNA expression cassettes were inserted into the deleted E1 region. The expression of spCas9-HF1 is driven by a tetracycline repressor-controlled CMV promoter comprising two binding sites for the repressor (2×TetO2). The expression of the individual targeting or non-targeting gRNAs is under control of a constitutive human U6 (hU6) promoter. The structure and sequence of the gRNAs are exemplarily shown for E1A gRNA 9 bound to its target site. The control vector containing only the Cas9 expression cassette is also depicted. (B) Target sequences for the individual gRNAs and their positions within the HAdV-5 genome (AY339865.1).

Article Snippet: Linear DNA fragments (gBLOCKS) containing individual targeting or non-targeting gRNAs under control of a human U6 promoter were generated by gene synthesis (Integrated DNA Technologies, Coralville, IA, USA), and the individual expression cassettes were inserted into the Hind II and NotI sites of the SpCas9-HF1 expression cassette-containing intermediate vector, giving rise to vectors pENTR-CMV-TetO2-spCas9-HF1-hU6-gRNA 1–10, containing a single targeting gRNA each, and to the control vectors containing non-targeting gRNAs (pENTR-CMV-TetO2-spCas9-HF1-hU6-NT gRNA) or containing only the spCas9-HF1 expression cassette (pENTR-CMV-TetO2-spCas9-HF1).

Techniques: Recombinant, Plasmid Preparation, Construct, Derivative Assay, Expressing, Binding Assay, Control, Sequencing

Journal: Oncotarget

Article Title: Dietary NiCl₂ causes G₂/M cell cycle arrest in the broiler's kidney.

doi: 10.18632/oncotarget.5934

Figure Lengend Snippet: Figure 3: Changes of p-ATM, p53, p21, p-Chk1, p-Chk2 and p-cdc25C protein expression levels in the kidney at 42 days of age. (Immunohistochemistry, ×400).

Article Snippet: Table 1: Antibodies used in immunohitochemistry Name Company Cat# Dilution p-ATM Bioss, China bs-2272R 1:100 p-Chk1 Bioss, China bs-5251R 1:100 p-Chk2 Bioss, China bs-3721R 1:100 p53 Boster, China BM0101 1:100 p21 Boster, China BA0272 1:100 p-cdc25C Bioss, China bs-3482R 1:100 p-cdc2 Boster, China BM0027 1:100 cyclinB1 Bioss, China bs-0572R 1:100 PCNA Boster, China BM0104 1:100 Oncotarget35969www.impactjournals.com/oncotarget or P < 0.01) in the 900 mg/kg groups at 14 days of age and in the three NiCl2-treated groups from 28 to 42 days of age in comparison with those in the control group.

Techniques: Expressing, Immunohistochemistry

Journal: Oncotarget

Article Title: Dietary NiCl₂ causes G₂/M cell cycle arrest in the broiler's kidney.

doi: 10.18632/oncotarget.5934

Figure Lengend Snippet: Figure 5: Changes of the mean density of p-ATM, p53, p21, p-Chk1, p-Chk2 and p-cdc25C protein expression in the kidney. Data are presented with the mean ± standard deviation (n=5×5) *P<0.05, compared with the control group **P<0.01, compared with the control group.

Article Snippet: Table 1: Antibodies used in immunohitochemistry Name Company Cat# Dilution p-ATM Bioss, China bs-2272R 1:100 p-Chk1 Bioss, China bs-5251R 1:100 p-Chk2 Bioss, China bs-3721R 1:100 p53 Boster, China BM0101 1:100 p21 Boster, China BA0272 1:100 p-cdc25C Bioss, China bs-3482R 1:100 p-cdc2 Boster, China BM0027 1:100 cyclinB1 Bioss, China bs-0572R 1:100 PCNA Boster, China BM0104 1:100 Oncotarget35969www.impactjournals.com/oncotarget or P < 0.01) in the 900 mg/kg groups at 14 days of age and in the three NiCl2-treated groups from 28 to 42 days of age in comparison with those in the control group.

Techniques: Expressing, Standard Deviation, Control

Journal: Oncotarget

Article Title: Dietary NiCl₂ causes G₂/M cell cycle arrest in the broiler's kidney.

doi: 10.18632/oncotarget.5934

Figure Lengend Snippet: Figure 7: Changes of ATM, p53, p21, Chk1, Chk2 and cdc25 mRNA expression levels in the kidney. Data are presented with the mean ± standard deviation (n=5) *P<0.05, compared with the control group **P<0.01, compared with the control group.

Article Snippet: Table 1: Antibodies used in immunohitochemistry Name Company Cat# Dilution p-ATM Bioss, China bs-2272R 1:100 p-Chk1 Bioss, China bs-5251R 1:100 p-Chk2 Bioss, China bs-3721R 1:100 p53 Boster, China BM0101 1:100 p21 Boster, China BA0272 1:100 p-cdc25C Bioss, China bs-3482R 1:100 p-cdc2 Boster, China BM0027 1:100 cyclinB1 Bioss, China bs-0572R 1:100 PCNA Boster, China BM0104 1:100 Oncotarget35969www.impactjournals.com/oncotarget or P < 0.01) in the 900 mg/kg groups at 14 days of age and in the three NiCl2-treated groups from 28 to 42 days of age in comparison with those in the control group.

Techniques: Expressing, Standard Deviation, Control

Journal: Journal of visualized experiments : JoVE

Article Title: Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity

doi: 10.3791/62182

Figure Lengend Snippet: Outlines PCR primer sequences used for the detection of select genes involved in neonatal keratinocyte plasticity.

Article Snippet: Use GAPDH as an internal control ( ). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Name of Material/ Equipment Company Catalog Number Comments/Description MCDB 153-LB basal media Sigma-Aldrich M7403 MCDB 153-LB basal media w/ HEPES buffer KSFM-scm ThermoFisher Scientific 17005042 Supplemented with 1% Penicillin/Streptomycin, 20 ng/ml EGF, 10 ng/ml basic fibroblast growth factor, 0.4% bovine serum albumin (BSA), and 4 μg/ml insulin Human Basic Fibroblast Growth Factor (hFGF basic/FGF2) Cell Signaling Technology 8910 Human Epidermal Growth Factor (hEGF) Cell Signaling Technology 8916 Human Insulin Millipore Sigma 9011-M Thermo Scientific™ Sterile Single Use Vacuum Filter Units Thermo Scientific 09-740-63D FITC-conjugated anti-integrinα6 Abcam ab30496 PE-conjugated anti-EGFR ( San Jose, CA; catalog number ) BD Pharmingen 555997 Dispase Sigma-Aldrich D4818 NEST Scientific 1-Well Cell Culture Chamber Slide, BLACK Walls on Glass Slide, 6/PK, 12/CS Stellar Scientific NST230111 P63 Thermo Scientific 703809 1:200 dilution Cytokeratin 14 Santa Cruz Biotechnology sc-53253 1:200 dilution All Prep DNA/RNA Mini Kit Qiagen 80204 iScript cDNA Synthesis Kit Bio-Rad 1708890 iQ SYBR Green Supermix (Bio-Rad) Bio-Rad 1708880 Promega TransFast kit Promega E2431 pMSCV-IRES-EGFP plasmid vector Addgene 20672 Open in a separate window Outlines PCR primer sequences used for the detection of select genes involved in neonatal keratinocyte plasticity.

Techniques: Cell Culture, SYBR Green Assay, Plasmid Preparation