1：Experimental Design and Method Selection:

The study designed folate-targeted lipid-polymer hybrid nanoparticles (TOI_HNPs) using a combination of two-step method and solution evaporation technique. Methods included formulation of NPs, characterization (size, zeta potential, morphology, stability, encapsulation efficiency), optical properties assessment, oxygen and ICG release studies, dual-mode imaging (ultrasound and photoacoustic), cellular uptake (confocal microscopy and flow cytometry), cytotoxicity (MTT assay), apoptosis (flow cytometry), ROS generation (fluorescence microscopy and microplate reader), and protein expression (Western blot).

2：Sample Selection and Data Sources:

Human SKOV3 ovarian cancer cells (FR positive) and A549 lung cancer cells (FR negative) were used. Cells were cultured in RPMI 1640 or DMEM with 10% FBS.

3：List of Experimental Equipment and Materials:

Equipment included rotary evaporator, ultrasonic probe, magnetic stirrer, centrifuge, confocal laser scanning microscope, transmission electron microscope, dynamic light scattering instrument, UV-Vis spectrophotometer, fluorescence spectrometer, dissolved oxygen meter, ultrasonic diagnostic instrument, photoacoustic imaging system, flow cytometer, fluorescence microplate reader, and Western blot apparatus. Materials included ICG, PLGA, lipids (DPPC, DPPG, DSPE-PEG-FA, cholesterol), PFP, PVA, SOSG, antibodies, dyes (DiI, DAPI, MTT, DCFH-DA), and other reagents.

4：Experimental Procedures and Operational Workflow:

NPs were formulated by forming a lipid film, mixing PFP with ICG solution, adding PLGA solution, sonicating, adding lipid film, evaporating solvent, and centrifuging. Characterization involved TEM, DLS, optical microscopy, UV-Vis, fluorescence spectroscopy, oxygen release, ICG release, dual-mode imaging in agar-gel phantom, cellular uptake in SKOV3 and A549 cells, cytotoxicity and apoptosis assays after laser and US exposure, ROS measurement, and Western blot for HIF-1α and IL-

5：Data Analysis Methods:

Data were analyzed using statistical methods (Student's t-test) with GraphPad Prism 6.0, with P<0.05 considered significant. Semi-quantitative analysis of imaging data used specific software.