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. 2023 Mar 14;12(9):10979–10989. doi: 10.1002/cam4.5781

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© 2023 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Effect of freezing rate on HCT116 phosphoproteomic profile. Effect of freezing rate on the phosphoproteome of cancer cell line HCT116. HCT116 samples were frozen in polypropylene vials in either liquid nitrogen (−196°C, in 2 s, N = 2; one sample was lost due to a technical error in the mass spectrometer) or in 25 s using pre‐cooled isopentane (−80°C, N = 3). (A) Unsupervised cluster analysis of all identified phosphopeptides does not separate samples with highest versus lowest freezing rate. (B) Venn diagram of overlapping phosphopeptides between all five samples shows reproducible phosphopeptide identification regardless of freezing rate. (C) Correlation by Pearson's r shows high correlation between phosphoproteomic profiles of samples frozen at high versus lower rate.