Ubiquitous DNase activity in invadosome-forming cells. (a–e) RAW264.7, THP-1, fish macrophages, MDA-MB-231, and MTC, with the former three forming podosomes and the latter two forming invadopodia, exhibited DNase activity in punctate patterns. (f) CHO-K1 cell as a control does not form invadosomes (no actin core), and no DNase activity was observed at the cell–substrate interface. (g) Percentage of actin cores that are colocalized with strong SNS punctate signal (20 cells for each data point, with 20–35 invadosomes analyzed for each cell). Error bars represent standard errors. (h) F-actin and SNS signal reporting DNase activity was coimaged in RAW264.7 cells transfected with LifeAct-GFP. (i) Time-series images of F-actin and SNS signal in one representative podosome (Video 1). The grid size is 2 × 2 µm². (j) Temporal curves of F-actin and DNase activity averaged over 20 podosomes, showing that DNase activity manifests in ∼1 min after actin core nucleation. Scale bars, 5 µm.