Structural investigation of C6/36 and Vero cell cultures infected with a Brazilian Zika virus - Fig 6

<p><b>C6/36 cells infected with ZIKV analyzed by transmission electron microscopy (TEM) at different time points post-infection (A-B: 48 hr p.i., C-D: 72 hr p.i.).</b> Several large viroplasm-like perinuclear compartments (V) (A-B) and ZIKV particles (*) measuring approximately 40–50 nm in diameter in the endoplasmic reticulum cisternae (RER) (A, B) and in lysosomes (L) (C-D) were observed. Nucleocapsids were observed inside the rER (D). Thickening of the nuclear membrane and rough endoplasmic reticulum cisternae (rER) (black head arrow) (C), numerous lysosomes (L) (C, D) and vesicular compartments associated with rER (arrow) (C) measuring approximately 100 nm in diameter were observed. Nucleus (N).</p

Uninfected C6/36 cells (negative controls) at different time points in culture.

<p><b>No cellular ultrastructural alterations were observed.</b> (A–B) 24 hr of culture; (C) 48 hr of culture; (D) 72 hr of culture. Rough endoplasmic reticulum cisternae (rER), nucleus (N).</p

Detection of ZIKV RNA and viral load quantification from C6/36 and Vero cell supernatants at 24, 48 and 72 hr p.i. by quantitative real-time RT-PCR.

<p>Detection of ZIKV RNA and viral load quantification from C6/36 and Vero cell supernatants at 24, 48 and 72 hr p.i. by quantitative real-time RT-PCR.</p

Immunolocalization of ZIKV E protein (4G2, green) [arrow] in the cytoplasm of Vero cells infected with ZIKV at different time points post-infection.

<p>(A) Uninfected cells; (B) 24 hr p.i; (C) 48 hr p.i.; (D) 72 hr p.i. The nuclei were counterstained with DAPI (blue). (E) The graph represents the mean ± standard deviation of the infection percentage (%). A higher number of infected cells was observed at 24 hr p.i.</p

Uninfected Vero cells (negative controls) at different culture time points.

<p>No cellular ultrastructural alterations were observed. (A) 24 hr of culture; (B) 48 hr of cultures; (C): 72 hr of culture. Nucleus (N), mitochondria (M).</p

Vero cell monolayer infected with ZIKV analyzed by phase contrast light microscopy at different time points post infection.

<p>(A) Uninfected cells with no monolayer changes; (B) 24 hr p.i.; (C) 48 hr p.i.; (D) 72 hr p.i. Cell individualization, rounding and detachment as well as cytolysis were observed at all time points and were more evident at 72 hr p.i.</p

Immunolocalization of ZIKV E protein (4G2, green) [arrow] inside cytoplasm of C6/36 cells infected with ZIKV at different time points post infection.

<p>(A) Uninfected cells; (B) 24 hr p.i.;(C/D) 48 hr p. i.; (E) 72 hr p.i. The nuclei (N) were counterstained with DAPI (blue). (F) The graph represents the mean ± standard deviation of the infection percentage (%). The highest number of infected cells was observed at 72 hr p.i.</p