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Lower expression of Gal-1 is correlated with higher viral loads produced by DENV-1-infected cells.

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posted on 13.11.2014, 03:37 by Karina Alves Toledo, Marise Lopes Fermino, Camillo del Cistia Andrade, Thalita Bachelli Riul, Renata Tomé Alves, Vanessa Danielle Menjon Muller, Raquel Rinaldi Russo, Sean R. Stowell, Richard D. Cummings, Victor Hugo Aquino, Marcelo Dias-Baruffi

(A) Gal-1 expression on HMVEC-L, Vero-E6 and ECV-304 cells was assessed by western blot method and normalized by β-actin endogenous control. The relative density of Gal-1 was determined by ImageJ software. (B) HMVEC-L, Vero-E6 and ECV-304 cells (2.5x104) were incubated with DENV-1 (MOI 0.5) for 72 hours at 37°C. At the end of incubation period, the total amounts of viral RNA in the cell-free supernatants were determined by Real-Time PCR, using a standard curve constructed from DENV-1 RNA purified from 1x107 PFU (PFU: plate formed units). Results are shown as Viral RNA amount equivalents to PFU/ml±SD from 3 independent assays performed in triplicates. (C) ECV-304 cells were inoculated with DENV-1 (MOI 0.5) or only with medium and cultivated for 72 hours at 37°C. Cells were analyzed for Gal-1 expression by western blot assay. (D) Soluble Gal-1 was detected in the supernatants from cell cultures using ELISA method (N = 3). *p<0.01; **p<0.001.