Journal: PLoS ONE

Article Title: Comparative characterization of flavivirus production in two cell lines: Human hepatoma-derived Huh7.5.1-8 and African green monkey kidney-derived Vero

doi: 10.1371/journal.pone.0232274

Figure Lengend Snippet: Cells were infected with JEV at MOI 0.1 under high confluency conditions as described in the legend to , and the culture supernatants were harvested at the indicated times. RNA was extracted and purified from each culture supernatant, and viral RNA copy number in the supernatant was determined by qRT-PCR and regarded as the number of virus particles. A virus titer in PFU of each culture supernatant was also determined by plaque assay, and particle-to-PFU ratio was calculated. Each symbol represents the relative value calculated by dividing the value of Huh7.5.1-8-derived JEV by that of Vero-derived JEV obtained from the same experiment. Bars with error bars represent the mean ± SD of the ratio from eight (1 and 2 d pi) or seven (3 d pi) independent experiments (the reason for the different sample sizes was that one experiment lacked data for 3 d pi). Statistical significance was determined by a one-sample t test. Values in parentheses indicate p values, and those less than 0.05 were considered statistically significant.

Article Snippet: Total RNA was extracted and purified from cells and culture supernatant using the Blood/Cultured Cell Total RNA Mini Kit (Favorgen Biotech, Pingtung City, Taiwan; No. FABRK 001–2) and the Viral Nucleic Acid Extraction Kit I (Favorgen Biotech; No. FAVNK 001–2), respectively, according to the manufacturer’s instructions.

Techniques: Infection, Purification, Quantitative RT-PCR, Plaque Assay, Derivative Assay

Journal: PLoS ONE

Article Title: Comparative characterization of flavivirus production in two cell lines: Human hepatoma-derived Huh7.5.1-8 and African green monkey kidney-derived Vero

doi: 10.1371/journal.pone.0232274

Figure Lengend Snippet: Cells were infected with JEV at MOI 0.1 under high confluency conditions as described in the legend to . Cells and culture supernatants were harvested at 0, 6, 9, 12, 24, 48, and 73 h pi, and RNA was purified. Viral RNA copy number in each RNA fraction was determined by qRT-PCR. The copy number of intracellular viral RNA was normalized by that of GAPDH RNA in the same fraction, whereas the amount of extracellular viral RNA was expressed as the total copy number in the culture supernatant of each well. The relative amounts of intracellular (A) and extracellular (B) viral RNA were calculated by dividing each value by that of Vero cells at 0 h pi in the same experiment. Data represent the mean ± SD of the relative values from four independent experiments. Graph (a) is an enlarged view of 0−12 h pi drawn on a linear scale, whereas graph (b) is a whole view drawn on a log scale. Some error bars are not visible due to their small size. Statistical significance was determined by a one-sample t test (0 h pi) or an unpaired two-tailed t test (other time points). Values in parentheses indicate p values, and those less than 0.05 were considered statistically significant. Huh, Huh7.5.1–8 cells.

Article Snippet: Total RNA was extracted and purified from cells and culture supernatant using the Blood/Cultured Cell Total RNA Mini Kit (Favorgen Biotech, Pingtung City, Taiwan; No. FABRK 001–2) and the Viral Nucleic Acid Extraction Kit I (Favorgen Biotech; No. FAVNK 001–2), respectively, according to the manufacturer’s instructions.

Techniques: Infection, Purification, Quantitative RT-PCR, Two Tailed Test

Journal: PLoS ONE

Article Title: Comparative characterization of flavivirus production in two cell lines: Human hepatoma-derived Huh7.5.1-8 and African green monkey kidney-derived Vero

doi: 10.1371/journal.pone.0232274

Figure Lengend Snippet: Cells were infected with JEV at MOI 0.1 under high confluency conditions as described in the legend to . Cells and culture supernatant were harvested at the indicated times. RNA was extracted and purified, and the viral RNA copy number in each RNA fraction was determined by qRT-PCR. The extracellular amount of viral RNA was expressed as a percentage of the total (extracellular plus intracellular) amount of viral RNA. Data represent the mean ± SD of three independent experiments. Each symbol represents the mean from one experiment. Statistical significance was determined by an unpaired two-tailed t test. Values in parentheses indicate p values, and those less than 0.05 were considered statistically significant.

Article Snippet: Total RNA was extracted and purified from cells and culture supernatant using the Blood/Cultured Cell Total RNA Mini Kit (Favorgen Biotech, Pingtung City, Taiwan; No. FABRK 001–2) and the Viral Nucleic Acid Extraction Kit I (Favorgen Biotech; No. FAVNK 001–2), respectively, according to the manufacturer’s instructions.

Techniques: Infection, Purification, Quantitative RT-PCR, Two Tailed Test