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Human cell F-actin density differentially influences trogocytosis and phagocytosis by Entamoeba histolytica Entamoeba histolytica is a parasitic amoeba and the cause amoebiasis, a common but understudied human diarrheal disease. E. histolytica trophozoites ("amoebae") kill human cells through a process of cell-nibbling called trogocytosis (trogo-: nibble) that contributes to tissue damage. Amoebae can also perform phagocytosis, in which entire human cells are ingested. Based on studies in which human cells were artificially stiffened, it was suggested that amoebae perform phagocytosis on stiffer cells, and trogocytosis on less stiff cells. A handful of recent studies of macrophages that used artificial targets or artificially stiffened target cells also suggested a similar relationship between target stiffness and trogocytosis/phagocytosis efficiencies. To better evaluate the impact of target cell stiffness on amoebic ingestion, instead of using artificial targets or artificial cell stiffening, we created human cell mutants in which individual Rho-pathway genes were knocked down. Strikingly, amoebae performed quantitatively reduced levels of trogocytosis on all knockdown mutants, regardless of cytoskeletal F-actin organization. In contrast, amoebic phagocytosis efficiency was inversely correlated with human cell cortical F-actin density. Thus, human cell F-actin organization differentially influences amoebic trogocytosis and phagocytosis. This is more complex than the conclusions of studies that used artificial targets or artificially stiffened cells. Our results emphasize that the dynamic nature of the cytoskeleton in living cells impacts trogocytosis. In addition to shedding light on the burgeoning field of eukaryotic trogocytosis, this work extends knowledge of amoebic ingestion processes that contribute to disease. ### Competing Interest Statement The authors have declared no competing interest. National Institute of Allergy and Infectious Diseases, R01AI146914, R21AI193905 https://www.biorxiv.org/content/10.64898/2026.03.17.712427v1