1：Experimental Design and Method Selection:

The study employed a reprecipitation method to synthesize carrier-free curcumin nanodrugs (Cur NDs) without toxic solvents. Methods included characterization of optical and physicochemical properties, drug release studies, cellular uptake analysis, cytotoxicity assays, ROS generation measurement, apoptosis assessment via flow cytometry, and Western blotting to investigate molecular pathways.

2：Sample Selection and Data Sources:

4T1 mouse breast cancer cells were used, purchased from ATCC. Curcumin (purity>94%) and other reagents were sourced from Sigma-Aldrich, GIBCO, and other suppliers.

3：List of Experimental Equipment and Materials:

Equipment included transmission electron microscope (Hitachi H-800), scanning electron microscope (Hitachi S4800), DLS instrument (Malvern Zetasizer Nano ZS), fluorescence spectrophotometer (Shimadzu RF-5301 PC), UV-vis spectrophotometer (Shimadzu 3100), inverted fluorescence microscope (IX71; Olympus), flow cytometer (FACS, Becton Dickinson), laser (LSR450SD-2W, LASEVER), multiwell plate reader (Bio Tek), and Western blot detection system (Tanon 4200). Materials included curcumin, ethyl alcohol, deionized water, RPMI-1640 medium, FBS, SRB, ROS kit, Annexin V-APC apoptosis kit, antibodies (JNK, p-JNK, Bax, caspase-3, GAPDH).

4：0). Materials included curcumin, ethyl alcohol, deionized water, RPMI-1640 medium, FBS, SRB, ROS kit, Annexin V-APC apoptosis kit, antibodies (JNK, p-JNK, Bax, caspase-3, GAPDH). Experimental Procedures and Operational Workflow:

4. Experimental Procedures and Operational Workflow: Cur NDs were synthesized by injecting curcumin in ethanol into water under stirring, followed by purification and lyophilization. Characterization involved TEM, SEM, DLS, UV-vis, and fluorescence spectroscopy. Drug release was studied using dialysis bags in PBS with/without light irradiation. Cellular uptake was assessed via fluorescence microscopy and flow cytometry after incubation with Cur NDs. Cytotoxicity was evaluated using SRB assay and flow cytometry after light exposure (450 nm, 640 mW, 1 min). ROS generation was measured using a fluorescence kit. Apoptosis and protein expression were analyzed via flow cytometry and Western blotting.

5：Data Analysis Methods:

Data were analyzed using GraphPad Prism for IC50 calculation, Student's t-test for two-group comparisons, and one-way ANOVA with Bonferroni's post hoc test for multiple groups. Protein band gray values were analyzed with Quantity One software.