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Compared to the emerging mechanisms underlying the strongly adhesive mesenchymal durotaxis, little is known about whether immune cells - migrating in amoeboid mode - could follow mechanical cues. Here, we develop an imaging-based confined migration device with a stiffness gradient. By tracking live cell trajectory and analyzing the directionality of T cells and neutrophils, we observe that amoeboid cells can durotax. We further delineate the underlying mechanism to involve non-muscle myosin IIA (NMIIA) polarization towards the soft-matrix-side but may not require differential actin flow up- or down-stiffness gradient. Using the protista\n                    <jats:italic>Dictyostelium<\/jats:italic>\n                    , we demonstrate the evolutionary conservation of amoeboid durotaxis. Finally, these experimental phenomena are theoretically captured by an active gel model capable of mechanosensing. Collectively, these results may shed new lights on immune surveillance and recently identified confined migration of cancer cells, within the mechanically inhomogeneous tumor microenvironment or the inflamed fibrotic tissues.\n                  <\/jats:p>","DOI":"10.7554\/elife.96821","type":"journal-article","created":{"date-parts":[[2024,5,24]],"date-time":"2024-05-24T11:26:11Z","timestamp":1716549971000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.96821","source":"Crossref","is-referenced-by-count":2,"title":["Amoeboid cells undergo durotaxis with soft end polarized NMIIA"],"prefix":"10.7554","volume":"13","author":[{"given":"Chenlu","family":"Kang","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02v51f717","id-type":"ROR","asserted-by":"publisher"}],"name":"Institute of Systems Biomedicine, School of Basic Medical Sciences, Peking University Health Science Center","place":["Beijing, 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