Extended Data Fig. 1: Dual tumour spatial distribution of NP1.

a,b, Representative TEM images of NP1 (a) and Ctrl-NP1 (b). Scale bars, 100 nm. c,d, Zeta-potentials recorded for NP1 (c) and Ctrl-NP1 (d). e,f, Size changes of NP1 (e) and Ctrl-NP1 (f) dispersed in water, PBS, or DMEM medium supplemented with 10% FBS at 4 °C at different times. g-i, CLSM images of MDA-MB-231 cells treated with NP1 (g) or Ctrl-NP1 (h,i) for 1 h. Scale bars, 10 μm. j-l, CLSM images of MDA-MB-231 cells after incubation with fresh medium containing 10% FBS and NP1 for 1 h. Scale bars, 20 μm in (j,k) and 50 μm in (l). m, CLSM images of MDA-MB-231 cells after incubation with NP1’ foe 1 h. Scale bar, 10 μm. The boxes shown the fields of (1 and 2) coexistence of cells and TEVs. n, CLSM images of MDA-MB-231 cells after incubation with NP1’ for 5, 15, 30, 60, or 180 min. Scale bar, 20 μm. o, Z-stack images of MDA-MB-231 cells after incubation with NP1’ for 1 h. Scale bar, 20 μm. p, Number-weighted size distribution of SEVs isolated from MDA-MB-231 cells by NTA after incubation with NP1 (SEVs^NP1), Ctrl-NP1 (SEVs^Ctrl-NP1), or PBS (SEVs) for 1 h, followed by fresh medium for 8 h. q,r, The fluorescence spectra (q) and quantitative analysis (r) of SEVs^NP1, SEVs^Ctrl-NP1, and SEVs. s, Number-weighted size distribution of LEVs isolated from MDA-MB-231 cells by NTA after incubation with NP1 (LEVs^NP1), Ctrl-NP1 (LEVs^Ctrl-NP1), or PBS (LEVs) for 1 h, followed by fresh medium for 8 h. t,u, The fluorescence spectra (t) and quantitative analysis (u) of LEVs^NP1, LEVs^Ctrl-NP1, and LEVs. v,w, Representative confocal fluorescence images for observing the colocalization of NP1 (red) with TEVs markers CD63 (green) (v) and TSG101 (green) (w), respectively. MDA-MB-231 cells were treated with NP1 for 1 h. The cell nuclei were stained with DAPI and are shown in blue. Scale bars, 5 μm. x, Excitation and emission fluorescence spectra of NP1^BODIPY (NP1’) and NP1^RhB (NP1). y, Time dependent fluorescence spectra change recorded for NP1^BODIPY/RhB dispersed in PBS at 37 °C. z, FRET ratio (I₅₈₀/I₅₃₈) of NP1^BODIPY/RhB in PBS at 37 °C. aa, CLSM images of MDA-MB-231 cells after incubation with NP1^BODIPY/RhB for 1 h. Scale bars, 20 μm. In all experiments, the concentration of NPs was 0.1 g/L. In e,f,r,u,z, n = 3 biologically independent samples. Data are presented as the mean ± s.d. Statistical significance was calculated via unpaired two-tailed Student’s t-test (r,u). In a-aa, experiments were repeated three times independently with similar results; representative images are shown in a,b,g-o,v,w,aa.

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