Fig 1.
EBER in situ hybridization (EBER-ISH) using commercially labelled probes and detection reagents.
Signals are detected as a purple colour and nuclei are countrerstained red. A) shows an example of TSCC where most cells are negative and a few tumour cells show faint nuclear staining (arrows). In tumour B) there is a predominantly cytoplasmic staining pattern in the majority of tumour cells, with some nuclear staining also seen. C) An example of a TSCC showing strong signals in the cytoplasm of tumour cells, but without nuclear staining.
Fig 2.
EBER in situ hybridization (EBER-ISH) using in-house prepared antisense and sense probes.
Signals are detected with DAB (brown) and nuclei are counterstained with haematoxylin (blue). A) Strong nuclear signal characteristic of EBER-ISH staining in tonsil from a patient with infectious mononucleosis used as a positive control. B) EBER-ISH of TSCC showing diffuse and speckled nuclear staining. C) Similar signals were seen using non-complementary sense probes.
Fig 3.
PCR for HBB and EBV BamH1 W fragment.
A 30 cycle PCR on 100 ng of gDNA showing amplification of HBB at 104 bp. HBB could not be amplified in samples 1 and 5. No amplification of EBV BamH1 W was seen in any sample. M = 100bp DNA marker; +C = positive control (100ng DNA extracted from the EBV-positive B95-8 cell line); -C = negative control (nuclease-free water).
Fig 4.
The presence of EBNA1 is detected with DAB (brown) and nuclei are counterstained with haematoxylin (blue). A) Positive control (EBV infected rabbit spleen) shows widespread nuclear staining. B) and C) show images of two representative TSCCs.