Real-time quantitative PCR analysis: RNA extraction

The MCF-7 breast cancer and HF cells at a density of 1×10⁵ cells/mL were seeded into 6-well plates and incubated for 24 hours. Afterward, the cells were treated by plasma, iron oxide NPs, and plasma–iron NPs. Then, the cells were cultured for 24 hours, and the total cellular RNA was extracted from treated and untreated cells using Trizol reagent (easy-BLUE Total RNA Extraction Solution) purchased from Intron Biotechnology, Korea, according to the manufacturer’s protocol (INTRON, cat17061). cDNA synthesis was performed using Revert Aid TM First-Strand cDNA Synthesis Kit (ABI, cat4368814) according to the manufacturer’s instruction (Thermo Fisher Scientific).