Figure 1.

ZIKV induces autophagy in HUVEC. (A) Confocal microscopy. HUVEC were infected with lentivirus expressing mTagRFP-mWasabi-LC3 followed by treatment at 48 h post-infection with mock treatment as a negative control, rapamycin (100 nM) treatment as a positive control, or infection of ZIKV (MOI = 1) for 24 h. The cells were observed by confocal microscopy with scale bars indicating 10 µm; (B) percent of cells with LC3 dots from total LC3-expressing cells was calculated. Data were derived from at least 100 cells for each sample. * p < 0.05, ** p < 0.01; (C) western blot analysis. The turnover of LC3-I to LC3-II was detected for mock-treated cells, rapamycin-treated (100 nM) cells, or the infected cells with ZIKV at MOI 1. Cells were harvested at indicated time points and detected with anti-LC3B antibody. GAPDH was used as a loading control. Representative images are presented; (D) the intensity band ratio of LC3-II to GAPDH. Graphs represent the ratio of LC3-II to GAPDH, as measured with densitometry; (E) HUVEC were infected with ZIKV at a range of MOIs (0.5, 1, and 2.5) for the indicated time, and LC3 expression was measured by western blot. The ratios of LC3-II to GAPDH were calculated and presented (F). Data are presented as means from three independent experiments. Significance was analyzed with two-tailed Student’s t test. * p < 0.05, ** p < 0.01.