Additional file 1 of Targeting and sensitizing MDR cancer by an MMP2 and pH dual-responsive ZnO-based nanomedicine
Creators: name: Zhou, Qing
name: Zhang, Li
name: Li, Yujiao
name: Wang, Jiao
name: He, Xiaolu
name: Zhang, Jieyu
name: Qiao, Youbei
name: Wu, Hong
name: Zhu, Lin
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Space Science
subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Biological Sciences not elsewhere classified
subject: Cancer
subject: Plasma Physics

description: Additional file 1. Fig. S1. Cellular uptake of the DOX or ZnO/DOX in the DOX-sensitive cancer cells and DOX-resistant MDR cancer cells after 1 h incubation, determined by flow cytometry. Mean fluorescence intensity of the flow cytometry data. Fig. S2. Confocal microscopic images of cellular internalization of the DOX or ZnO/DOX in the MDA-MB-231 and NCI/ADR-RES cells after 1 h incubation. Cell nuclei were stained by Hoechst. The scale bar is 100 μm. Fig. S3. Uptake of the DOX, ZnO/DOX, or ZnO + DOX mixture after 1 h incubation with the MDR cancer cells, determined by flow cytometry. MFI, mean fluorescence intensity. Fig. S4. Cytotoxicity of the DOX, ZnO NPs, and ZnO/DOX in DOX-sensitive cancer cells and DOX-resistant MDR cancer cells. Cell incubation time: 24 h. Fig. S5. Mitochondrial membrane potential after 1 h or 24 h treatments, determined by the JC-1 assay. The decrease in the JC-1 aggregate/monomer ratio indicates the mitochondrial depolarization. Confocal microscopic images of the JC-1 staining after 4 h treatments. The scale bar is 50 μm. Fig. S6. The C/EBP Homologous Protein levels after 24 h treatments, determined by a CHOP ELISA kit. Cell line: NCI/ADR-RES cells. Fig. S7. Drug penetration through the MDA-MB-231 and NCI/ADR-RES cell spheroids after 4 h incubation, determined by confocal microscopy. The scale bar is 200 μm. The right panel is the curves of the normalized mean fluorescence intensity vs the distance from the spheroid bottom to the top. Fig. S8. Synthetic scheme of the PEG-pp-PE.TLC chromatograms of the PEG-pp and PEG-pp-PE.1H NMR spectrum of the PEG-pp-PE. Fig. S9. Mean particle sizes of the ZnO/DPPG/PEG-pp-PE/DOX upon the incubation with HBSS or HBSS containing 10% FBS for 72 h. Particle sizes were measured by dynamic light scattering. Fig. S10. Digital photographs of the ZnO/DPPG/PEG-PE/DOX and ZnO/DPPG/PEG-pp-PE/DOX with the MMP2 incubation for 12 h. Particle sizes of the ZnO/DPPG/PEG-pp-PE/DOX with/without the MMP2 pretreatment, determined by dynamic light scattering. Fig. S11. Cellular uptake of the DOX-loaded ZnO-based NPs in the MDR cancer cells after 1 h incubation, determined by flow cytometry. MFI, mean fluorescence intensity. Confocal microscopic images of the cellular internalization of ZnO/DPPG/PEG-PE/DOX and ZnO/DPPG/PEG-pp-PE/DOX with or without the MMP2 pretreatment after 1 h incubation with the MES-SA/Dx5 cells. Cell nuclei were stained by Hoechst. The scale bar is 100 μm. Data were expressed as the mean ± SD, **p &lt; 0.01; ***p &lt; 0.001. Fig. S12. Morphology of the NCI/ADR-RES cell spheroids after treatments. The scale bar is 200 μm. Fig. S13. Cytotoxicity of the DOX-loaded ZnO-based NPs with/without the MMP2 pretreatment after 24 h incubation with the MDR cancer cells. Data were expressed as the mean ± SD, **p &lt; 0.01; ***p &lt; 0.001.