Additional file 1 of Self-assembly of DNA nanostructure containing cell-specific aptamer as a precise drug delivery system for cancer therapy in non-small cell lung cancer
Creators: name: Wang, Ning
name: Yu, Chang
name: Xu, Tingting
name: Yao, Dan
name: Zhu, Lingye
name: Shen, Zhifa
name: Huang, Xiaoying
Publisher: name: figshare
Year: 2022
Subjects: Biophysics
Biochemistry
Space Science
Medicine
Cell Biology
Molecular Biology
Pharmacology
Biotechnology
Chemical Sciences not elsewhere classified
Cancer

description: Additional file 1: Scheme S1. The self-assembly process from single-strand DNA to a structural unit. The detail base sequences of three single-strand DNA, including S6 (aptamer), PP (palindrome probe), and Linker, are displayed in the pattern diagram. Figure S1. 8% PAGE image showing the influence of palindrome sequences on the synthesis of DNA nanostructure. CP (common probe) was instead of PP (palindrome probe). C-NS represented the synthetic products from self-assembly system in the absence of palindrome sequences. Figure S2. The calculation of Apt-NS sizes according to AFM images. Frequency distributions of the length and width of Apt-NS were assessed using NanoScope analysis software based on a method reported in the literature [1]. The average length and width of Apt-NS were 101 ± 30 nm and 58 ± 8 nm, respectively. Figure S3. Stability assay of Apt-NS. (A) Serum stability assay of Apt-NS. PAGE analysis of samples such as Linker, PP, and Apt-NS (left panel). Quantitative assessment of Apt-NS serum stability based on gel electrophoresis images (right panel). (B) DNase I stability assay of Apt-NS. PAGE analysis of Apt-NS samples (upper panel). Quantitative assessment of Apt-NS DNase I stability based on gel electrophoresis images (lower panel). Figure S4. Assessment of cell recognition ability of the different concentrations of S6 aptamer on A549 cells. (A) Flow cytometric assay and (B) CLSM images showing the internalization of the different concentrations of S6 aptamer into A549 cells. Scale bar is 25 μm. Figure S5. Digital photos of DOX-loaded Apt-NS. (A, B) Photographs of free DOX (i) and DOX-loaded Apt-NS solution (ii) before and after centrifugation. Figure S6. Confocal images showing the time-dependent cellular uptake of DOX and Apt-NS-DOX by HELF cells. (A) Cellular uptake of free DOX by HELF cells. (B) Cellular uptake of Apt-NS-DOX by HELF cells. Red fluorescence indicated the DOX, green fluorescence indicated Cy5 labeled nanomaterials. Scale bar is 25 μm. Figure S7. Schematic illustration of the internalization and degradation of Apt-NS-DOX in A549 cells. Internalized Apt-NS-DOX was transferred to lysosomes, and then the DOX was released by lysosomes degradation. The released DOX could enter into nucleus and induced DNA damage. Figure S8. Apoptosis effects of Apt-NS-DOX on HELF cells. (A) Flow apoptosis assay of HELF cells treated with Apt-NS, DOX, and Apt-NS-DOX, respectively (Q1 means dead cells; Q2 means late apoptotic cells; Q3 means early apoptotic cells; Q4 means normal cells). (B) Quantitative analysis of cell apoptosis rate of each group in (A). Statistical analysis: ***P &lt; 0.001 and ns, no significance. Figure S9. Nanomaterial biodistribution and tumor-targeting imaging in vivo. (A) fluorescence images of the major organs (heart, liver, spleen, lung and kidney) and tumor tissues. (B) Quantitative analysis of the average fluorescence intensities of each group in (A). Statistical analysis: **P &lt; 0.01. Figure S10. Hemolysis assay for drug delivery system (Apt-NS-DOX). (+) represented a positive control (H2O) and (-) represented a negative control (PBS). Inset: the photographs of RBCs after incubation with the corresponding samples. The value presented as mean ± SD (n = 3). Figure S11. The calculation of Ki67-positive cells percentages according to the immunohistochemical staining of Ki67 in tumor tissues sections of different treatment groups. The value presented as mean ± SD (n = 5). Statistical analysis: ***P &lt; 0.001. Figure S12. The calculation of apoptotic cell percentages according to the TUNEL staining of tumor tissues sections in different treatment groups. The value presented as mean ± SD (n = 3). Statistical analysis: ***P &lt; 0.001. Table S1. Sequences of ssDNA designed in our experiments.