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Figure 4 | Molecular Cancer

Figure 4

From: Hepatitis B virus X protein specially regulates the sialyl lewis a synthesis among glycosylation events for metastasis

Figure 4

The inhibition of SLA expression in HBx-transfected cells and HBx-tranfected cell adhesion to endothelial cells by ST3Gal III, FUT III and FUT VII siRNAs. (A) Each cell was co-transfected with ST3Gal III, FUT III and VII siRNAs, as well as negative control siRNA, respectively. Total RNA from these cells was isolated. The mRNA expression of ST3Gal III, FUT III and VII were detected by RT-PCR, using primers which are indicated in Table 1. β-actin was included as an internal control. (B) To check whether SLA expression induced in HBx-transfected cells was suppressed by the siRNAs against ST3Gal III, FUT III and FUT VII genes, the expression was examined using flow cytometric analysis and immunofluorescent staining. Fluorescence histograms of a representative experiment indicate the baseline expression of SLA. The y-axis gives the percentage of positive cells (counts); the x-axis gives fluorescence intensity. (C) Each cell was co-transfected with ST3Gal III, FUT III and VII siRNAs and negative control siRNA (N.C.), respectively. Each cell was added on TNF-α-stimulated HUVECs, and was incubated for 20 min at 37°C with rotation at 60 rpm in a shaking incubator. The attached cells were visualized using a microscope, as described in Materials and Methods.

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