Additional file 1 of Combination of polythyleneimine regulating autophagy prodrug and Mdr1 siRNA for tumor multidrug resistance
Creators: name: Wang, Changduo
name: Li, Zhipeng
name: Xu, Ping
name: Xu, Lisa
name: Han, Shangcong
name: Sun, Yong
Publisher: name: figshare
Year: 2022
Subjects: subject: Biophysics
subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Biological Sciences not elsewhere classified
subject: Cancer
subject: Hematology
subject: Infectious Diseases
subject: FOS: Health sciences

description: Additional file 1:Figure S1. The synthetic route of PEI-PTX polymers. (a) succinyloxide, pyridine, 25 ℃; (b) HATU, HOBt, DIPEA, 25 ℃. Figure S2. FT-IR spectrums of PTX and PTX-SA. Figure S3. 1H NMR spectrum of PTX (A) and PTX-SA (B) in DMSO-d6 (400 MHz). Figure S4. Mass spectrum of PTX-SA. HRMS (ESI): exact mass calculated for [M + K]+ (C51H55NO16) requires m/z 976.56, found m/z 976.72. Figure S5. FT-IR spectrums of PTX-SA, PEI, PEI-PTX, and simple mixed of PEI &amp; PTX-SA. Figure S6. 1H NMR spectrum of PTX, PEI-PTX, and PEI (400 MHz, DMSO-d6). Figure S7. Gel Permeation Chromatography spectrums of PTX, PEI, and PEI-PTX. Figure S8. Determination of the critical micelle concentration (CMC) of PEI-PTX. Figure S9. The Entrapment efficiency of FAM-siRNA at various weight ratios of PEI-PTX and FAM-siRNA by fluorophotometer. Figure S10. Pictures of PP, PP/siRNA, and PP/siRNA/HA nanoassembles in PBS. Figure S11. (A) The siRNA protection of nanoassembles in 50% FBS at various times (0-24 h). (B) The heparin resistance ability of nanoassembles at various ratios (heparin/siRNA, IU/μg). Figure S12. Pictures of hemolytic toxicity study with different concentrations (50, 100, 250, 500, and 1000 μg/mL) of PP or PP/HA, 0.9% NaCl as negative control, and TritonX-100 as positive control. Figure S13. (A) The accumulative release of PTX from PP under different conditions of pH5.0, pH7.4, pH5.0 + Esterase, and pH7.4 + Esterase (n=3). (B) The accumulative release of PTX from PP/siRNA/HA under different conditions of pH5.0, pH7.4, and pH5.0 + HAase (n=3, *P＜0.05, ***P＜0.001). Figure S14. (A) Flow cytometry analysis of internalization in A549 cells after treated with naked FAM-siRNA, PP/FAM-siRNA or PP/FAM-siRNA/HA for 1h and 4 h. (B) Flow cytometry analysis of internalization in A549/T cells (n=3, ***P＜0.001). Fig. S15 (A) The Endo/lysosomal escape of PP/FAM-siRNA/HA in A549 cells by CLSM at 1 h or 4 h after 2 h uptake. Lysosomal were stained by LysoTracker (red), colocalization analysis of FAM-siRNA and lysosome at 1 h (B) or 4 h (C) by imageJ in A549 cells. (D) The Endo/lysosomal escape of PP/FAM-siRNA/HA in A549/T cells by CLSM at 1 h or 4 h after 2 h uptake. Lysosomal were stained by LysoTracker (red). Colocalization analysis of FAM-siRNA and lysosome at 1 h (E) or 4 h (F) in A549/T cells. Figure S16. Cytotoxicities of PEI in A549 and A549/T cells. Cell viability treated with various concentrations of 1.8K PEI. Figure S17. Cell apoptosis study of PTX, PP, PP/siRNA, PP/HA, and PP/siRNA/HA towards A549 cells (A) and A549/T cells (B). Figure S18. The cell-cycle distribution of A549 cells (A) and A549/T cells (B), and quantitive presentation of cell-cycle (C). Figure S19. H&amp;E staining of tumor tissues.