MCF-7 cells were seeded in a 48-well plate at a density of ∼ 2 × 104 cells per well 24 hours a day before the experiment. At the day of incubation, cells were washed three times with PBS, and as-synthesized 2-nm Au-POY2T NPs and self-assembled nanoflowers (calculated as the number of Au-POY2T NPs from the gold element amount) were added at a concentration of 100 nM and incubated at different times (1, 3, 6, 12, and 24 hours) at 37°C. After each incubation, the cells were washed three times with PBS buffer. Cell lysis buffer (250 μl per well) was used to lyse the cells, and the culture plate was kept at room temperature on a vibrator for 30 min. In addition, a freeze/thaw cycle was adopted to further facilitate the cell lysis process before the ICP sample preparation step.

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