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IHC / IMMUNOFLUORESCENCE
ISH / FISH
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CONTROL TMA
An Example of Pathology-Model Based Target Validation
Background: An in vitro study had suggested that molecule X secreted by tumor cells might induce T cell apoptosis, so as to help tumor cells escape from T cell mediated tumor cell killing. The purpose of this project was to find evidence supporting this suggestion.
Procedures
Results
Picture
Antibody production
Specific antibody was produced. The antibody recognized a single band with correct molecular weight in Western blot (as indicated by the arrow).
IHC (left) and TUNEL (right) analyses to correlate apoptic index with expression levels of the molecule in our apoptosis related model.
High expression level of the molecule (left top brown staining indicated by the arrow) did not correlate with the apoptotic index in the model. The brown dots on the right indicated by arrows are apoptotic bodies.
IHC detection of CD4 and CD8 populations in the apoptosis model.
There were abundant CD4 and CD8 lymphocytes (indicated by arrows) near or within the tumor cells that showed high level of molecule X.
Conclusion
Our pathology model-based analyses showed that high level of molecule X in the tumors did not correlate with high apoptotic index among tumor infiltrating T cells, directly against the original hypothesis that molecule X might induce T cell apoptosis.