Quantifying Cellular Protein Activation

Immunohistochemistry was performed as previously described [45 (link)]. The following antibodies were used: phospho-STAT3 Y705, phospho-Histone H2AX S139, phospho-Histone H3 S10, and cleaved Caspase 3 (all from Cell Signaling Technology, Danvers, MA). After image acquisition using an Aperio Scanscope XT (Leica), the staining indexes were calculated dividing the number of positive epithelial cells/nuclei by the total number of epithelial cells/nuclei, using Aperio Imagescope software (Leica Biosystems). The resulted ratio is shown as index.

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De Oliveira T., Ramakrishnan M., Diamanti M.A., Ziegler P.K., Brombacher F, & Greten F.R. (2018). Loss of Stat6 affects chromatin condensation in intestinal epithelial cells causing diverse outcome in murine models of inflammation-associated and sporadic colon carcinogenesis. Oncogene, 38(11), 1787-1801.

Publication 2018

phospho-STAT3 Y705 phospho-Histone H2AX S139 phospho-Histone H3 S10 cleaved Caspase 3 Aperio Scanscope XT Aperio Imagescope software

After image Antibodies Caspase 3 Histone h2ax Histone h3 Immunohistochemistry Nuclei Stat3

Corresponding Organization : German Cancer Research Center

Other organizations : Georg Speyer Haus, International Centre for Genetic Engineering and Biotechnology, University of Cape Town

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Variable analysis

independent variables

Phospho-STAT3 Y705
Phospho-Histone H2AX S139
Phospho-Histone H3 S10
Cleaved Caspase 3

dependent variables

Staining indexes
Number of positive epithelial cells/nuclei
Total number of epithelial cells/nuclei

control variables

Aperio Scanscope XT (Leica) for image acquisition
Aperio Imagescope software (Leica Biosystems) for data analysis

controls

No positive or negative controls were explicitly mentioned.

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