Additional file 1 of The double-edged sword effect of macrophage targeting delivery system in different macrophage subsets related diseases
Creators: name: Yuan, Yuchuan
name: Long, Ling
name: Liu, Jiaxing
name: Lin, Yongyao
name: Peng, Cuiping
name: Tang, Yue
name: Zhou, Xuemei
name: Li, Shuhui
name: Zhang, Chengyuan
name: Li, Xiaohui
name: Zhou, Xing
Publisher: name: figshare
Year: 2020
Subjects: subject: Space Science
subject: Medicine
subject: Sociology
subject: FOS: Sociology
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified

description: Additional file 1: Fig. S1. Characterization of nanoparticles. (a–c) Representative TEM images of PS (a), COOH-PS (b) and DEX-PS (c). (d) Infrared spectra analysis of various PS. (e) Representative fluorescent images of various PS. (f) Size distribution of PS determined by DLS. (g) Zeta potential of PS. Scale bars in (a–c) mean 500 nm. Data in (g) are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S2. (a) Cell toxicity of PS, COOH-PS and DEX-PS to Raw 264.7 cells for 24 h. (b) Fluorescence intensity of PS, COOH-PS and DEX-PS at 0.01 mg/mL determined by a fluorescence spectrophotometer. (c) The uptake rates of various nanoparticles by Raw 264.7 cells at various time points. Data are presented as the mean ± SD. n = 3. Fig. S3. (a, b) Cell surface expression of the M1 marker (iNOS and CD86) and the M2 marker (MMR and CD163) as analyzed by flow cytometry. (c) Representative phase contrast images of Raw 264.7 cells and polarized macrophages, bar indicated 20 μm (d) Release of TNF-α, IL-1β, IL-10 and TGF-β in cell culture media was analyzed by using ELISA. (e–f) The mRNA level of pro-inflammatory cytokines (e) and anti-inflammatory cytokines (f) expressed by various cells determined by PCR. Data are presented as the mean ± SD. n = 3. Fig. S4. The mean fluorescence of whole cells in BMDM-M1 cells (a) and in BMDM-M2 cells (b) after incubation with various NPs. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S5. The expression changes of MMR on M1 macrophages after incubating with nanoparticles for 4 h. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S6. (a, b) The uptake rate of PS (a) and COOH-PS (b) by M1 macrophages influenced by various inhibitors. (c, d) The uptake rate of PS (c) and COOHPS (d) by M2 macrophages influenced by various inhibitors. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S7. (a) The percent of Nile Red+ cells in MMR+ and MMR− cells. (b) The fluorescence intensity of MMR+ and MMR− cells. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S8. (a) Quantification illustrating distribution of Nile Red fluorescence signals in isolated whole blood. (b) Fluorescence intensity of eight hours after injection of PSs nanoparticles in vivo. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S9. (a) Representative confocal laser-scanning microscopy images of the M1 marker (iNOS) and M2 marker (CD163) expression in 4T1 tumor. (b) The ratio of CD86 (M1 marker)/CD206 (M2 marker) in acute peritonitis induced by zymosan. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S10. (a) Quantitative statistical results of NPs distribution in tumor. Data are presented as the mean ± SD (n = 2 in saline group, while n = 3 in other groups). (b) Quantitative statistical results of NPs distribution in acute peritonitis lesion. Data are presented as the mean ± SD (n = 3). (c) The ex vivo fluorescent images of heart, liver, lung, kidney and spleen. (d) Quantitative statistical results of NPs distribution in major organs. Data are presented as the mean ± SD (n = 3), * means p &lt; 0.05. Fig. S11. The percent of CD11b+ B220− cells in Nile Red+ blood cells. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S12. The uptake rate of PS (a) and COOH-PS (b) by Raw 264.7 cells influenced by various inhibitors. Data are presented as the mean ± SD. n = 3, * means p &lt; 0.05. Fig. S13. The expression changes of CD86 on M1 macrophages after treated with various nanoparticles. Data are presented as the mean ± SD. n = 3. Fig. S14. The survival time of mice in zymosan induced acute peritonitis. Data are presented as the mean ± SD. n = 10, * means p &lt; 0.05. Fig. S15. Growth of subcutaneous 4T1 tumors after treated with various PS. Data are presented as the mean ± SD. n = 5. Fig. S16. The distribution of nanoparticles in tumor observed by CLSM.