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Any peptides defined as false positive by the analysis of the buffer slides were excluded, and the remaining peptide responses were normalized using the model Yijk = slidei+subarrj+blockk where Yijk is the response (i.e. the index) from block k in sub-array j on slide i.
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The Coble creep and GB sliding are excluded for dominant deformation mechanism.
After rinsing with buffer, slides were dehydrated in ethanol.
Slides with <100 cells were excluded and an average of 223 cells per slide was scored (range 100 452).
For negative control DAB was excluded from the reaction buffer.
Following a series of washes in borate buffer, the slides were mounted in borate buffer for immediate imaging.
After rinsing in wash buffer, the slides were incubated with HRP conjugated appropriate secondary antibody.
Still in the citrate buffer the slides were cooled at room temperature for 20 min.
Citrate buffer was pre-heated and slides were subsequently added for 15 min (microwave 300 W).
Antigen retrieval was performed in buffer citrate (pH 6.0) and slides were microwaved.
Slides were removed from labelling buffer and excess buffer surrounding the tissue was wiped away.
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