Journal: Molecular Cancer
Article Title: ZEB1 limits adenoviral infectability by transcriptionally repressing the Coxsackie virus and Adenovirus Receptor
Figure Lengend Snippet: Up-regulation of cell-surface CAR levels upon ZEB1 knockdown coincides with increased adenoviral infectability . A and B . Cell-surface CAR levels following ZEB1 knockdown were measured by flow cytometry on PANC-1 cells treated with TGF-β1 ( A ), or on MDA-MB-231 cells ( B ). Error bars represent standard deviations (biological duplicates). C and D . Ad-GFP uptake following ZEB1 knockdown was determined by flow cytometry measuring GFP levels ( C ), or by real-time PCR for virus copy number ( D ). Error bars represent standard deviations. A-D . CAR and GFP signals are expressed as geometric means of the fluorescence signal intensities normalized to the untreated controls (A-C), virus copy numbers as relative adenovirus fiber levels (D). Abbreviations: UT, untransfected; Ctrl #1, siControl ON-TARGETplus Non-targeting siRNA #1 (Dharmacon); Ctrl #2, firefly luciferase-targeting siRNA; ZEB1 siRNA #1/#2, ZEB1-targeting siRNAs. Ctrl #2 and ZEB1 siRNA sequences are provided in Additional file 1 (Table S3). p
Article Snippet: Flow cytometry Live cells were stained with an anti-CAR-phycoerythrin (PE) antibody (E1-1, mouse monoclonal; Santa Cruz Biotechnology, Inc.) or PE-conjugated control IgG-PE (mouse monoclonal IgG1 κ, Pharmingen/BD Biosciences) while rotating for 60 minutes at 4°C.
Techniques: Flow Cytometry, Cytometry, Multiple Displacement Amplification, Real-time Polymerase Chain Reaction, Fluorescence, Luciferase