1：Experimental Design and Method Selection:

The study involved preparing BSA-coated gold nanorods (AuNRs) via a seed-mediated growth method, characterizing them, and using them for photothermal ablation of breast tumor cells (4T1-Luc). Immune responses were assessed by co-culturing ablated cells with immature dendritic cells (DCs) using direct contact and diffusion models to distinguish effects of cell-cell interaction and soluble factors.

2：Sample Selection and Data Sources:

Mouse breast tumor cells (4T1-Luc) and mouse bone marrow-derived immature DCs were used. Cells were cultured in specific media (RPMI1640 for tumor cells, MEM-α for DCs) with supplements like FBS and GM-CSF.

3：List of Experimental Equipment and Materials:

Equipment included TEM (JEM-2100F), zeta-potential analyzer (ELSZ-2000), FTIR spectrometer (8400S), UV-VIS spectrophotometer (V-660), ICP-OES (Leeman Prodigy), laser (805 nm, Thorlabs), digital thermometer, microplate reader (Benchmark Plus), fluorescence microscope (Olympus), and ELISA kits. Materials included HAuCl4, NaBH4, CTAB, AgNO3, ascorbic acid, BSA, WST-1 assay kit, calcein-AM/PI staining kit, and various chemicals and cell culture reagents.

4：Experimental Procedures and Operational Workflow:

AuNRs were synthesized, coated with BSA, and characterized. Photothermal performance was tested with laser irradiation. Cell viability and uptake of AuNRs were assessed. Tumor cells were ablated with laser, and DCs were co-cultured with ablated cells in different models (direct contact, transwell for diffusion). Cytokine secretion (IL-6, IL-12, IL-1β, TNF-α) from DCs was measured via ELISA after 24 hours.

5：Data Analysis Methods:

Data were analyzed using one-way ANOVA with significance at p < 0.05. Results are presented as mean ± standard deviation from triplicate experiments.