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Fig. 2 | Virology Journal

Fig. 2

From: Evaluation of the virucidal effects of rosmarinic acid against enterovirus 71 infection via in vitro and in vivo study

Fig. 2

Time-of-addition analysis of rosmarinic acid (RA) against EV71 infection. a Viral VP1 mRNA was detected by quantitative real-time polymerase chain reaction. RD cells were treated with 10 μM RA before (− 2 h), during (0 h), or post (+ 2 h) EV71 (MOI = 1; 6 × 104 PFU) infection for 1 h, then viral RNA was extracted. b Viable viruses were detected by plaque reduction assay. RD cells were treated with 10 μM RA before (− 2 h), during (0 h), or post (+ 2 h) EV71 (MOI = 1; 6 × 104 PFU) infection for 1 h, then viable viruses were detected by plaque reduction assay. c Plaque reduction assay. EV71 (6 × 104 PFU) was mixed with 0, 1, 3, 5, or 10 μM RA for 1 h at 37 °C. Virus-treated and mock control tubes were diluted 20-fold and applied to infect RD cells for 1 h. d Immunofluorescence assay. RD cells (1 × 104) were infected with EV71 at a MOI of 1 (1 × 104 PFU) and treated simultaneously with or without 10 μM RA at 37 °C. At 1 h post infection, images were obtained with a fluorescence microscope (100× magnification). Green fluorescence was produced by Alexa Fluor 488-FITC and blue fluorescence was produced by DAPI. Each assay was repeated three times. Data were obtained from three separate experiments and are presented as the mean ± S.D. *P < 0.05, **P < 0.001, as compared with control

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