Journal: Journal of Virology

Article Title: Extracellular Conformational Changes in the Capsid of Human Papillomaviruses Contribute to Asynchronous Uptake into Host Cells

doi: 10.1128/JVI.02106-17

Figure Lengend Snippet: Infectious internalization of FPC-HPV16 is faster and more synchronous than that of HPV16. (A) In an add-on experiment, HPV16 (dotted) or FPC-HPV16 (solid) was used to infect HeLa (left) or HaCaT (right) cells. At indicated times postinfection, extracellular virus was inactivated by a high-pH wash (pH 10.5). Depicted are the half-times of infectious internalization and the difference in the half-time between the two viruses in hours and percentages. (B) A seed-over experiment was performed as described for panel A. Infectious internalization values were normalized to the 48-h samples and are depicted in percentages ± standard errors of the means (SEM) for all experiments. Curves were fitted with the nonlinear regression function of GraphPad Prism v6. Statistical significance was tested by two-tailed unpaired Student's t test in GraphPad Prism v6; P values are indicated by asterisks: *, P < 0.05; **, P < 0.01. (C) pHrodo-HPV16 and pHrodo-FPC-HPV16 particles were added to cells, and the pHrodo signal was imaged at different time points p.i. and analyzed as relative intensity per cell.

Article Snippet: Statistical significance was tested by two-tailed unpaired Student's t test in GraphPad Prism v6; P values are indicated by asterisks: *, P < 0.05; **, P < 0.01. (C) pHrodo-HPV16 and pHrodo-FPC-HPV16 particles were added to cells, and the pHrodo signal was imaged at different time points p.i. and analyzed as relative intensity per cell.

Techniques: Two Tailed Test

Journal: Journal of Virology

Article Title: Extracellular Conformational Changes in the Capsid of Human Papillomaviruses Contribute to Asynchronous Uptake into Host Cells

doi: 10.1128/JVI.02106-17

Figure Lengend Snippet: Infectious internalization kinetics of HPV16 are unaffected by cell cycle synchronization. (A) Schematic depiction of cell cycle synchronization by double thymidine block. (B) Synchronized and nonsynchronized cells were fixed with ethyl alcohol (EtOH) and analyzed for their cell cycles states at different times postrelease. Cell cycle phases were designated according to cellular DNA content by PI staining and flow-cytometric analysis. G1, a single set of chromosomes (i.e., DNA content = 1); G2/M, duplicated chromosomes (DNA content = 2); S, replicating chromosomes between the two states. Shown are the values from two independent experiments. (C) Infectious internalization of HPV16 (upper panels) and FPC-HPV16 (lower panels) with synchronized (black dotted line) and nonsynchronized (gray dotted line) cells was analyzed in add-on experiments. Cells were infected at 2 h, 6 h, or 14 h postrelease from thymidine block. Curves were fitted with the nonlinear regression function of GraphPad Prism v6. Relative infectious internalization values were normalized to 48-h infection samples and are depicted as percentages ± SEM.

Article Snippet: Statistical significance was tested by two-tailed unpaired Student's t test in GraphPad Prism v6; P values are indicated by asterisks: *, P < 0.05; **, P < 0.01. (C) pHrodo-HPV16 and pHrodo-FPC-HPV16 particles were added to cells, and the pHrodo signal was imaged at different time points p.i. and analyzed as relative intensity per cell.

Techniques: Blocking Assay, Staining, Infection