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Flow cytometry requires a suspension for staining and analysis.
Slides were randomly picked from each group for staining and analysis.
Embedded blocks were step serially cut to produce 7 micron sections for staining and analysis.
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See Additional file 1: Supplementary Information for more information about the protocols followed for antibody staining and analysis of MSI and gene mutations.
Subsequently, the supernatant was removed for analysis and the cells were washed, fixed, and permeabilized for intracellular staining and analysis by FACS using Cytofix/Cytoperm Plus kit as per manufacturer's instructions (Pharmingen).
The tissues were sectioned using microtome and were fixed on glass slides for further staining and analysis.
We then cut the brains into 50 μm coronal sections on a freezing microtome and took 1 4 sections for subsequent staining and analysis.
Frozen samples were then oriented and 6 µm thick cryostat sections were cut for immunohistochemical staining and analysis using light microscopy, as previously described (Di Stefano et al, 2002).
Brains were cut into 50 μm sagittal sections on a freezing microtome with 1 4 sections being taken for subsequent staining and analysis.
For IF staining and analysis of sub-cellular localization of C-YFP Bax and C-YFP BaxFP, the two genes were ando Cytned in-frame to a Flag tag or an HA tag in the pc N-YFPor pEGFP expression vec N-YFPrespectheely.
Tumors established following injection of SCC cells into both flanks of an FVB mouse were removed at day 7. Tumor tissue was processed to obtain single cell suspension for staining and subsequent FACS analysis (antibodies listed in Table S2).
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