Fig. 1

Experimental design and stable expression of the EBNA1 protein; (a) Overview of the strategy used to identify the changes in splicing in EBNA1-expressing cells. A high throughput RT-PCR approach was selected to look at the AS dynamics of numerous genes with a known involvement in cancer; (b) Western-Blot analysis showed expression of the EBNA1-HA-FLAG construct, as detected at 80- kDa by the anti-HA (1:1000), the anti-FLAG (1:1000) and the anti-EBNA1(1:100) antibodies upon stable selection of transfected HEK293T cells. Actin was used as a loading control on the same membrane after stripping. Each lane was charged with 20 μg of total protein. (c) Immunofluorescence of HEK293T-EBNA1-HA-FLAG cells using anti-HA antibody showed nuclear localization of the EBNA1-HA-FLAG protein, which confirm both the expression of the protein and its predicted cellular localization. Nuclei were stained using DAPI, and anti-HA and DAPI were images were merged. Scale bar, 10 μm