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. 2008 Oct 31;4(10):e1000189. doi: 10.1371/journal.ppat.1000189

Figure 4. Effect of HSPG-expression and HSPG-sulfation on Ad3 infection of CHO cells.

Figure 4

(A) Hexon staining. CHO-K1 cells were infected with Ad3 (MOI 512 pfu/cell). Three days post-infection, cells were fixed and stained for adenovirus hexon protein (red), E-cadherin as a cell surface marker (green), and nuclei (DAPI, blue). Magnification 40×. (B) MTT assay. CHO-K1, CHO-pgsA-745, and CHO-pgsE-606 cells were infected with various MOIs of Ad3 (0, 64, 128, 256, 512, 1024 pfu/cell). Seven days post-infection, mitochondrial activity of cells was determined via MTT assay. Data points represent the mean and standard deviation of experiments performed in triplicate. (C) CPE assay. CHO-K1, CHO-pgsA-745, and CHO-pgsE-606 cells were infected with Ad3 in a range of 0–2048 pfu/cell and monitored for CPE as described in Materials and Methods. Five days post-infection, cells were fixed and stained with crystal violet. Representative pictures are shown. Upper row: Photographs of crystal violet stained wells. Lower row: Photographs (Magnification 40×) of crystal violet stained wells. Presence of CPE is indicated with black borderlines. (D) Ad3 viral replication assay. Fold increase of Ad3 viral genomes 5 days post-infection of CHO-K1, CHO-pgsA-745, CHO-pgsE-606, and A549 cells is shown. Bars represent the mean and standard deviation of experiments performed in duplicate. (B–D) These experiments were independently repeated twice with similar outcomes.