Journal: International Journal of Molecular Sciences

Article Title: Roburic Acid as a Therapeutic Candidate: Antiproliferative Activity and Secondary Cell Death Response in Colorectal Cancer Cells

doi: 10.3390/ijms27052478

Figure Lengend Snippet: Representative images from the alkaline comet assay. DLD-1: ( A ) control, ( B ) RA IC 50 ; HCT-116: ( C ) control, ( D ) RA IC 50 ; HT-29: ( E ) control, ( F ) RA IC 50 ; and ( G ) bleomycin (20 μM)-treated cells used as a positive control. The images are shown at identical magnification (40×).

Article Snippet: Comet assay reagents: normal melting point agarose, low-melting point agarose, bleomycin sulfate, sodium hydroxide solution, EDTA, disodium salt dihydrate, tris(hydroxymethyl)aminomethane (TRIS), Triton X-100, sodium chloride solution, and 4,6-diamidino-2-phenylindole (DAPI) were purchased from Merck/Sigma Aldrich Chemical Co. (Burlington, MA, USA).

Techniques: Alkaline Single Cell Gel Electrophoresis, Control, Positive Control

Journal: Viruses

Article Title: Development of a Novel Method to Detect AAV Vector Integration

doi: 10.3390/v18030315

Figure Lengend Snippet: Generation and characterization of HeLa-AAV2-eGFP single-cell clones for AAV genome integration assay. ( A ) Schematic overview of the workflow used to establish HeLa-AAV2-eGFP single-cell clones. HeLa cells were treated with or without 50 uM of bleomycin for 2 h prior to AAV2-eGFP transduction at 50,000 vg/cell. Following 24 h of transduction and two weeks of culture, eGFP + cells were enriched by flow cytometry and expanded for four weeks. A second round of single-cell sorting was used to obtain individual eGFP + clones, which were cultured for an additional six weeks, after which the single-cell clones were analyzed by ( B ) percentage of eGFP expression by flow cytometry and ( C ) vector copy number by ddPCR using WPRE (red) and eGFP (green) primers and probes.

Article Snippet: Cells (1 × 10 5 per well) were seeded in 6-well plates, incubated overnight, and treated with or without 50 μM of bleomycin sulfate (Selleck Chemicals, S1214-10MG, Houston, TX, USA) for 2 h. After washing with DMEM containing 2% FBS, cells were transduced with AAV2-CAG-GFP particles (Addgene, 37825-AAV2, Watertown, MA, USA; 7 × 10 12 vector genomes per mL (vg/mL) at 5 × 10 4 vg/cell for 24 h).

Techniques: Single Cell, Clone Assay, Transduction, Flow Cytometry, Cell Culture, Expressing, Plasmid Preparation