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. 2021 Dec 10;12:766293. doi: 10.3389/fphar.2021.766293

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Copyright © 2021 Simpson, Roberts, Thompson, Leiper, Gittens, Trotter, Duckworth, Papoutsopoulou, Miyajima, Roberts, O’Kennedy, Rhodes and Campbell.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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C. difficile toxins A and B mediate increased Rac1 glucosylation within intestinal epithelial cells, which is unaffected by pre-treatment with soluble NSP from plantain. Immunoblot analysis shows an increase in Rac1 glucosylation following treatment with 10 ng/ml (A) toxin A (TcdA) for 24 h and (B) toxin B (TcdB) for 48 h, which is unaffected by pre-treatment of Caco-2 cells with 10 mg/ml soluble plantain NSP (P). Protein bands were analysed by chemiluminescence image densitometry. Mono-glucosylated Rac1 levels (assessed with antibody 102) were normalised to total Rac1 (assessed with antibody 238a), with representative blots shown for N = 3 experiments, n = 3 replicates. Significant differences from non-toxin treated controls, *p < 0.05, **p < 0.01 and ***p < 0.001; Kruskal-Wallis test.