ALT levels in the sera from mice at 7 dpi were measured using a Cobas C111 analyzer (Roche) in the NUS comparative medicine in-house veterinary diagnostic laboratory. Histopathological analysis was performed as previously described by Costa et al. (10 (link)). Briefly, liver samples from humice were obtained at day 7 after DENV-2 inoculation. Afterward, samples were immediately fixed in 10% buffered formalin for 24 h and embedded in paraffin. Tissue sections (4 mm thick) were stained with hematoxylin and eosin (H&E) and evaluated under an Axioskop 40 microscope (Carl Zeiss, Göttingen, Germany) adapted to a digital camera (PowerShot A620; Canon, Tokyo, Japan). Histopathology scoring was carried out by a pathologist in a blinded fashion according to a set of custom-designed criteria described by Costa et al. (10 (link)). Hepatocyte swelling, degenerative changes, necrosis, and hemorrhage were each graded on a scale of 0 to 5 (0, absent; 1, minimal; 2, slight; 3, moderate; 4, marked; and 5, severe). The sum of the score was computed to derive the overall liver damage score. A total of two sections for each animal were examined.
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