Journal: mBio

Article Title: Adaptation of Human Parainfluenza Virus to Airway Epithelium Reveals Fusion Properties Required for Growth in Host Tissue

doi: 10.1128/mBio.00137-12

Figure Lengend Snippet: HAE infection with HPIV3 HN H552Q: emergence of variants. (A) Timeline of viral growth after infection of HAE with wt HPIV3 (blue) and HPIV3 HN H552Q (red). (B) Plaque diameter in CV1 cells 16 to 18 h after infection with the indicated virus retrieved from HAE ( n = 20 per virus from 3 experiments). (C) Titers of viruses grown in HAE: wt HPIV3 (blue), HPIV3 HN H552Q (red), or HPIV3 “small plaques” (brown). Data points are means (±SD) of triplicate measurements and are representative of at least 5 experiments. (D) Quantitation of viral genome released 3 days post-HAE infection (determined by qRT-PCR). Data points are means (±SD) of triplicate experiments. (E) Percentage of infectious virions (out of total particles) retrieved from infection of HAE.

Article Snippet: To measure the amount of viral genome released, 30 μl of fluid containing released virus was processed for RNA extraction, and then the genome was quantified using the real-time one-step qRT-PCR pathogen detection kit specific for HPIV3 (PrimerDesign) as per the manufacturer’s instructions.

Techniques: Infection, Virus, Quantitation Assay, Quantitative RT-PCR

Journal: mBio

Article Title: Adaptation of Human Parainfluenza Virus to Airway Epithelium Reveals Fusion Properties Required for Growth in Host Tissue

doi: 10.1128/mBio.00137-12

Figure Lengend Snippet: Mutations acquired in HN and F glycoproteins of HPIV3 variants

Article Snippet: To measure the amount of viral genome released, 30 μl of fluid containing released virus was processed for RNA extraction, and then the genome was quantified using the real-time one-step qRT-PCR pathogen detection kit specific for HPIV3 (PrimerDesign) as per the manufacturer’s instructions.

Techniques: Variant Assay, Mutagenesis, Virus

Journal: mBio

Article Title: Adaptation of Human Parainfluenza Virus to Airway Epithelium Reveals Fusion Properties Required for Growth in Host Tissue

doi: 10.1128/mBio.00137-12

Figure Lengend Snippet: Molecular structure of wild-type and mutant HN and F. (A) HN dimer structure (PDB IV2I), with the monomers colored gray and cyan. The locations of the H552Q and Q559R mutations at the dimer interface are highlighted by showing residues H552 and Q559 as sticks and marked with asterisks. The structure was subjected to 20 ns of MD simulation. (B) Crystal structure of the postfusion HPIV3 F protein (right) and a homology model of the prefusion protein, based on PIV5 F (left). The three chains of F are colored green, cyan, or magenta. The G396 residues are represented as yellow spheres.

Article Snippet: To measure the amount of viral genome released, 30 μl of fluid containing released virus was processed for RNA extraction, and then the genome was quantified using the real-time one-step qRT-PCR pathogen detection kit specific for HPIV3 (PrimerDesign) as per the manufacturer’s instructions.

Techniques: Mutagenesis

Journal: mBio

Article Title: Adaptation of Human Parainfluenza Virus to Airway Epithelium Reveals Fusion Properties Required for Growth in Host Tissue

doi: 10.1128/mBio.00137-12

Figure Lengend Snippet: Characteristics of variant HPIV3 HN and F proteins

Article Snippet: To measure the amount of viral genome released, 30 μl of fluid containing released virus was processed for RNA extraction, and then the genome was quantified using the real-time one-step qRT-PCR pathogen detection kit specific for HPIV3 (PrimerDesign) as per the manufacturer’s instructions.

Techniques: Variant Assay, Binding Assay, Activation Assay