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While physical cues are increasingly recognized as coregulators of macrophage activation alongside biochemical signals, the role of pathogen-induced biophysical remodeling in shaping immune function remains poorly understood. In this study, we employed noncontact scanning ion conductance microscopy (SICM) to monitor real-time nanomechanical and morphological changes in immortalized bone marrow-derived macrophages (iBMDMs) at single-cell resolution during lipopolysaccharide (LPS)-induced M1 polarization. We found that M1 activation significantly increased the surface area-to-volume (SA\/V) ratio and decreased cellular stiffness\u2500biophysical adaptations that may support enhanced pro-inflammatory responses. Mechanistically, these changes were driven by NF-\u03baB\u2013dependent activation of the cytoskeletal regulator MARCKS. The inhibition of MARCKS disrupted both morphological remodeling and inflammatory activation, highlighting its essential role in coordinating immune signaling with nanomechanical plasticity. This study provides the first real-time, quantitative evidence linking inflammatory signaling to macrophage morphological and nanomechanical remodeling, and shows that MARCKS inhibition suppresses, while PMA-mediated activation mimics, LPS-induced responses.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c01115","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T10:06:43Z","timestamp":1785924403000},"page":"23475-23486","source":"Crossref","is-referenced-by-count":0,"title":["Real-Time Scanning\nIon Conductance Microscopy Reveals\nMARCKS-Driven Morphological Remodeling and Nanomechanical Softening\nduring LPS-Induced Macrophage Polarization"],"prefix":"10.1021","volume":"98","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2732-0808","authenticated-orcid":true,"given":"Dong","family":"Wang","sequence":"first","affiliation":[{"name":"Kanazawa University , , ,","place":["Kanazawa, Japan, 920-1192"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ritsuko","family":"Yamase","sequence":"additional","affiliation":[{"name":"Kanazawa University , , ,","place":["Kanazawa, Japan, 920-1192"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyunghee","family":"Park","sequence":"additional","affiliation":[{"name":"Kanazawa University , , ,","place":["Kanazawa, Japan, 920-1192"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiromi","family":"Nishikawa","sequence":"additional","affiliation":[{"name":"Kanazawa University , , ,","place":["Kanazawa, Japan, 920-1192"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1799-4625","authenticated-orcid":true,"given":"Andrew","family":"Shevchuk","sequence":"additional","affiliation":[{"name":"Kanazawa University , , ,","place":["Kanazawa, Japan, 920-1192"]},{"name":"Imperial College London , , ,","place":["London, U.K., W12 0NN"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7359-6023","authenticated-orcid":true,"given":"Rebecca C.","family":"Coll","sequence":"additional","affiliation":[{"name":"Queen\u2019s University Belfast , , ,","place":["Belfast, U.K., BT9 7BL"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuri","family":"Korchev","sequence":"additional","affiliation":[{"name":"Kanazawa University , , ,","place":["Kanazawa, Japan, 920-1192"]},{"name":"Imperial College London , , ,","place":["London, U.K., W12 0NN"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Wang","sequence":"additional","affiliation":[{"name":"Queen\u2019s University Belfast , , ,","place":["Belfast, U.K., BT9 7BL"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3123-1405","authenticated-orcid":true,"given":"Yanjun","family":"Zhang","sequence":"additional","affiliation":[{"name":"Kanazawa University , , ,","place":["Kanazawa, Japan, 920-1192"]},{"name":"Imperial College London , , ,","place":["London, U.K., W12 0NN"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"issue":"1","key":"2026081809085517800_cit1","doi-asserted-by":"publisher","first-page":"207","DOI":"10.1038\/s41392-023-01452-1","article-title":"Macrophages in immunoregulation\nand therapeutics","volume":"8","author":"Chen","year":"2023","journal-title":"Signal Transduction Targeted\nTher."},{"issue":"12","key":"2026081809085517800_cit2","doi-asserted-by":"publisher","DOI":"10.1002\/cti2.1222","article-title":"Origins\nand diversity of macrophages in health and disease","volume":"9","author":"Sreejit","year":"2020","journal-title":"Clin. 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