s typhimurium atcc 14028 cell morphology change  (ATCC)

Journal: Viruses

Article Title: CD46 Is a Protein Receptor for Human Adenovirus Type 64

doi: 10.3390/v16121827

Figure Lengend Snippet: Ad64.eGFP delivers GFP gene via CD46. ( A ) Pre-treatment of HeLa cells with 10 mU neuraminidase from Vibrio cholerae to remove cell surface sialic acid or 10 mM EGTA to chelate calcium ions prior to incubation with Ad64.eGFP significantly decreased susceptibility to viral entry as determined by cell fluorescence compared to uninfected cells (Analysis of variance followed by Tukey’s test (ANOVA-T), n = 3 for each treatment). Error bars represent standard deviations. ( B ) HAdV-B16 fiber knob (16 FK) blocked Ad5 pseudotyped with Ad16 fiber (Ad5.F16) and HAdV-D64 (Ad64), but not HAdV-C5 (Ad5) gene delivery into HeLa cells in a concentration-dependent manner. Experiments were performed in duplicate. ( C ) Anti-CD46 antibody blocked calcium-dependent entry into HeLa cells (ANOVA-T, n = 3 for each treatment). ( D ) Expression of CD46 BC1 or C1 isoform on CHO cells conferred susceptibility to Ad64.eGFP entry (ANOVA-T, n = 3 for each treatment). Single asterisk, p < 0.05; double asterisk, p < 0.0001.

Article Snippet: Previous molecular studies on HAdV-D37 employed Chang conjunctival cells [ , , , ], which were later shown to be contaminated with HeLa cells during establishment (ATCC), or A549 lung epithelial cells [ , ].

Techniques: Incubation, Fluorescence, Concentration Assay, Expressing

Journal: Viruses

Article Title: CD46 Is a Protein Receptor for Human Adenovirus Type 64

doi: 10.3390/v16121827

Figure Lengend Snippet: Direct binding of HAdV-D64 to soluble CD46. ( A ) Ad64.eGFP particles were co-precipitated with His-tagged soluble CD46 (sC) using Ni-NTA Agarose then subject to SDS-PAGE. Main panel lanes: 1, CD46 (~45–50 kDa) mixed with Ad64.eGFP; 2, flowthrough from NiNTA Agarose, 3–5 wash fractions, 6–8 elution fractions. Hx, HAdV-D64 hexon. Gel images of ultracentrifugation-purified Ad64.eGFP (top left) and purified sCD46-C (bottom left) are shown to the left for comparison. Masses of molecular standards are shown in kDa. ( B ) 5 nm Ni-NTA-NanoGold beads (Au) were incubated with CD46, Ad64.eGFP particles, or both, filtered through a 0.1 µm filter, visualized using transmission electron microscopy at 60,000× magnification, and counted. Only the presence of the large adenovirus particle and CD46 led to substantial retention of NanoGold beads in the filter (ANOVA-T, n = 3 for each treatment). Double asterisks, p < 0.0001.

Article Snippet: Previous molecular studies on HAdV-D37 employed Chang conjunctival cells [ , , , ], which were later shown to be contaminated with HeLa cells during establishment (ATCC), or A549 lung epithelial cells [ , ].

Techniques: Binding Assay, SDS Page, Purification, Comparison, Incubation, Transmission Assay, Electron Microscopy

Journal: Viruses

Article Title: CD46 Is a Protein Receptor for Human Adenovirus Type 64

doi: 10.3390/v16121827

Figure Lengend Snippet: HAdV-D64 can enter human conjunctival epithelial cells. ( A ) Pre-treatment of HCjE cells with soluble CD46-C (sCD46) and/or neuraminidase inhibited Ad64.eGFP viral entry, as determined by expression of green fluorescence of eGFP transgene (ANOVA-T, n = 5). ( B ) Ad64.eGFP vector entry into HCjE cells in Keratinocyte-SFM in the presence or absence of EGTA, a calcium chelator (ANOVA-T, n = 3). ** denotes p < 0.001.

Article Snippet: Previous molecular studies on HAdV-D37 employed Chang conjunctival cells [ , , , ], which were later shown to be contaminated with HeLa cells during establishment (ATCC), or A549 lung epithelial cells [ , ].

Techniques: Expressing, Fluorescence, Plasmid Preparation