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Table 1.

All antibodies used for detection in this study.

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Figure 1.

Phosphoprotein IHC scoring.

H scores of staining intensity for all studied biomarkers, by treatment condition. “2+2” is the rapid fixation condition, “24 hr.” is the standard 24-hour room temperature condition without pre-fixation cold ischemia, and “Ischemia” denotes a 1-hour cold ischemic period prior to 24-hour room temperature fixation. The medians are represented by horizontal lines, the boxes in plots correspond to the 25th and 75 percentiles of each distribution, the whiskers extend to the most extreme value that is within 1.5 times the distance between the first and third quartiles, and data beyond the whiskers are shown as dots.

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Table 2.

Comparison of Pathologist H-scoring between conditions for all tested biomarkers.

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Figure 2.

Colon Cancer Phosphoprotein IHC.

Representative results from IHC analysis of two colonic carcinoma samples subjected to either the rapid fixation condition (“2+2”) or standard 24-hour room temperature condition without pre-fixation cold ischemia (“24 hr”). pERK, pMTOR, pMEK1/2 and pPRAS40 are shown for one case and pBAD and pAKT are shown from a second case. Two different cases are shown here because not all cases showed staining for all markers. All micrographs are taken at 200× with equivalent exposures.

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Figure 3.

Colon Cancer Phosphoprotein IHC.

Representative images of one colonic carcinoma case stained with antibodies to BRAF V600E, pEGFR, EGFR, and pMSK1, showing significant loss of pEGFR staining in the ischemia condition. All micrographs are taken at 200× with equivalent exposures.

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Figure 4.

Colon Cancer Phosphoprotein IHC.

Representative of a second colonic carcinoma case stained with antibodies to BRAF V600E, pEGFR, PTEN, and pMSK1, showing significant loss of pEGFR staining in the ischemia condition. All micrographs are taken at 200× with equivalent exposures.

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