1：Experimental Design and Method Selection:

The study involved synthesizing nanoGold-core dendrimer nanoparticles (Au-DEN) by functionalizing gold nanoparticles with thiolated dendrons, conjugating methotrexate (MTX) for targeting and therapeutic effect, and loading IR780 for photothermal activity. Methods included citrate reduction for Au NP synthesis, Au-thiol chemistry for dendron conjugation, esterification for MTX conjugation, and dialysis for purification. In vitro assays (drug release, photothermal response, cell viability, cellular uptake, ROS generation, hemolytic toxicity, stability) were conducted to evaluate the NPs.

2：Sample Selection and Data Sources:

Mouse macrophage RAW264.7 cells and LPS-activated RAW264.7 cells were used for cellular studies. Blood from mice was used for hemolytic assays. Materials were sourced from commercial suppliers (e.g., Sigma Aldrich, Fisher Scientific).

3：7 cells and LPS-activated RAW7 cells were used for cellular studies. Blood from mice was used for hemolytic assays. Materials were sourced from commercial suppliers (e.g., Sigma Aldrich, Fisher Scientific). List of Experimental Equipment and Materials:

3. List of Experimental Equipment and Materials: Equipment included Zetasizer Nano ZS-90 for DLS, UV spectrophotometer (UV 1800, Shimadzu), HPLC system (Agilent), SEM (JEOL JSM7600F), confocal laser scanning microscope (Leica), fluorescence microscope (Zeiss), orbital shaker, ultracentrifuge (Sorvall WX 80+), lyophilizer (Il Shin biobase), NIR laser (808 nm wavelength). Materials included gold chloride, dendrons, MTX, IR780, EDC, DMAP, cell culture media, and buffers.

4：Experimental Procedures and Operational Workflow:

Synthesis of Au NPs, conjugation with dendrons to form Au-DEN, conjugation with MTX to form Au-DEN-MTX, loading of IR780 to form Au-DEN-MTX-IR

5：Characterization included size, PDI, zeta potential, UV-Vis spectra, FTIR, DSC, SEM. In vitro studies:

7 drug release in different pH buffers, photothermal response with NIR irradiation, cell viability (MTT assay), cellular uptake (fluorescence and CLSM), ROS generation (DCFH-DA assay), hemolytic toxicity, and stability studies at different temperatures.

6：Data Analysis Methods:

Data were analyzed using one-way ANOVA and two-way ANOVA with post-tests (Tukey-Kramer, Bonferroni) in GraphPad Prism software. Results are expressed as mean ± standard deviation.