BALB/c nude mice (6 weeks, female) were obtained from Beijing Charles River Laboratory Animal Technology Co. Ltd. (Beijing, China) and raised in a specific pathogen–free grade laboratory. All animal experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals proposed by the National Institutes of Health (NIH). All animal experiments were conducted with the approval of the Institutional Animal Care and Use Committee, Institute of Process Engineering, Chinese Academy of Sciences. Briefly, the MCF-7 tumor model was established by subcutaneous injection of 106 MCF-7 cells into the right flank of BALB/c nude mice (about 20 g) at day 0. After tumor volumes reached about 50 mm3, the animals were randomly divided into nine groups (four mice per group). Then, the mice were treated with 100 μl of various formulations (equivalent to 10 μM in POY2T sequence) at days 9, 12, and 15 by intravenous injection. In groups ③, ④, ⑤, and ⑥ (as labeled in Fig. 5), the tumors were irradiated with an 808-nm NIR laser (1 W/cm2) for 10 min (to reach the local temperature above 41°C) at 1, 3, 6, and 12 hours after each intravenous injection. In day 24, the mice were sacrificed, and all tumors were isolated and weighted.

Statistical analysis. All data are expressed as means ± SD. Statistical differences were determined by two-tailed Student’s t test; *P < 0.05, **P < 0.01, and ***P < 0.001. The density of Western blot bands was quantified using ImageJ software (NIH Image).

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