Additional file 1 of De novo design of a novel AIE fluorescent probe tailored to autophagy visualization via pH manipulation
Creators: name: Huang, Xueyan
name: Chen, Fei
name: Ma, Yeshuo
name: Zheng, Fan
name: Fang, Yanpeng
name: Feng, Bin
name: Huang, Shuai
name: Zeng, Hongliang
name: Zeng, Wenbin
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Space Science
subject: Cell Biology
subject: Neuroscience
subject: Physiology
subject: FOS: Biological sciences
subject: Pharmacology
subject: Biological Sciences not elsewhere classified
subject: Cancer

description: Additional file 1: Scheme S1. Synthetic routine of the ASMP-AP. Figure S1. The constructed potential energy scan surface of both S0 and S1 states of ASMP-AP as a function of O7-H49. Figure S2. Fluorescence spectra of ASMP-AP (10 µM) in different solvents. λex = 375 nm. Figure S3. Absorption spectra of ASMP-AP (10µM) in DMSO and in aqueous solution (1% DMSO). Figure S4. AIE properties of ASMP-AP. a) Fluorescence spectra of ASMP-AP (10 µM) in DMSO/water mixtures with different volume fraction of water (fw). b) Fluorescence emission at 560 nm of ASMP-AP (10 µM) in DMSO/water mixed solvent. λex = 375 nm. Figure S5. Absorption spectra of ASMP-AP in aqueous solution (10% DMSO) with different pH. Figure S6. DLS spectra of ASMP-AP (10 µM) in PBS buffer at different pH values. Figure S7. Fluorescence responses of ASMP-AP (10 µM) to pH 3.33 and 7.40 in the absence and presence of possible interfering substances at 37 °C, respectively. Such as metal ions (100 µM), essential thiols (1 mM), H2O2 (100 µM). Figure S8. Emission maximum versus switching cycles between pH 3.33 and pH 7.4. Figure S9. Cytotoxicity of ASMP-AP to Hela cells determined by MTT assay. The data are based on the average and show the standard deviation (n = 3). Figure S10. CLSM images of cells in in nutrient-rich medium at different time nodes. λex = 405 nm; λem = 560-620 nm. Scale bars, 10 μm. Figure S11. Autophagy-related protein expression and statistical analysis under starvation conditions. Figure S12. Lysosome-targeting properties of ASMP-AP in HeLa cells after 4 h-starvation treatment. Colocalization images stained with LysoTracker Green (1.0 μM, green channel, λex = 488 nm, λem = 500-550 nm) and ASMP-AP (10 μM, yellow channel, λex = 405 nm, λem = 560-620 nm), and the correlation of ASMP-AP and LysoTracker Green intensities as well as the intensity profiles within the ROI. Scale bars, 10 µm. Figure S13. Analysis of the fluorescence intensity of ASMP-AP channel in panel Figure 6A (n = 6, by ImageJ software). Statistical analyses were performed with One-way Anova. *: p &lt; 0.05, ****: p &lt; 0.0001. Figure S14. Confocal fluorescence images of zebrafish larvae in rapamycin induced autophagy. A) Followed by the pretreatment with rapamycin for 12 h at 28 ℃, zebrafish larvae were stained with ASMP-AP (10 μM, λex = 405 nm, λem = 560-620 nm) and LysoTracker green (1.0 μM, λex = 488 nm, λem = 500-550 nm). Statistical analyses were performed with One-way Anova. *: p &lt; 0.05, **: p &lt; 0.01, ***: p &lt; 0.001, and ****: p &lt; 0.0001. B) Colocalization relationship of ASMP-AP with LysoTracker green. Scale bars, 100 µm. Figure S15. Confocal fluorescence images of different positions of zebrafish larvae (eyes and midbrain) with rapamycin treatment. ASMP-AP (10 μM, λex = 405 nm, λem = 560-620 nm). Scale bars, 100 µm. Table S1. Comparison of properties of various pH-sensitive AIE probes. Figure S16. 1H NMR spectra (DMSO-d6)) of compound 1. Figure S17. 13C NMR spectra (DMSO-d6)) of compound 1. Figure S18. HRMS spectrum of compound 1. Figure S19. 1H-NMR spectra (DMSO-d6) of compound 2. Figure S20. 13C NMR spectra (DMSO-d6) of compound 2. Figure S21. HRMS spectrum of compound 2. Figure S22. 1H-NMR spectra (DMSO-d6) of compound 3. Figure S23. HRMS spectrum of compound 3. Figure S24. 1H NMR spectra (DMSO-d6) of compound 4. Figure S25. 13C NMR spectra (DMSO-d6) of compound 4. Figure S26. HRMS spectrum of compound 4. Figure S27. 1H NMR spectra (DMSO-d6) of ASMP-AP. Figure S28. 13C NMR spectra (DMSO- d6) of ASMP-AP. Figure S29. HRMS spectrum of compound ASMP-AP.