Curcumin

Curcumin (purity >98%; C805205) was purchased from Macklin (Shanghai, China). To prepare the oil solution, 0.16 g Curcumin, 0.8 g lecithin from soybean (Macklin, L812366), 1.2 g ethoxylated hydrogenated castor oil (Perfemiker, Shanghai, China, PA43980) and 2.5 g glyceryl tridodecanoate (Macklin, G810435) were mixed and stirred adequately. The appearance of the oil solution without Curcumin, Curcumin in saline, and prepared Curcumin oil solution is displayed in Figure 1B.

To determine the hydrophobicity of GDH, the ANS fluorescent probe method, SDS binding method, and curcumin were used in this study (Cardamone and Puri, 1992 (link); Kato et al., 2002 (link); Noh et al., 2006 (link); Sneharani, 2016 (link); Kamshad et al., 2019 (link)). ANS and curcumin were purchased from Macklin Biochemical Co., Ltd. (Shanghai, China). GDH hydrophobicity was assayed at 30°C, using a sample containing 20 mmol/L Tris–HCl buffer (pH 7.4), 500 mmol/L NaCl, and 0.6 g/L protein. Subsequently, 20 μl ANS (8 mmol/L) was added to 1 ml of the aforementioned sample. The mix was left in the dark for 30 min. The fluorescence intensity (λ_ex 390 nm and λ_em 504 nm) of the samples was monitored at 30°C after shocking. The concentration of pure protein ranged from 0 to 5 g/L. The curve slope represents the protein hydrophobicity index (S₀). For SDS binding and curcumin method, the absorbance of base blue SDS-methylene and fluorescence intensity of riboflavin were detected, which reflects the number of hydrophobic groups on the protein surface.

All oligonucleotides were synthesized and HPLC-purified by Sangon Biotechnology Co., Ltd. (Shanghai, China) and are listed in Table S6. Ammonium persulfate (APS), N,N,N,N-tetramethylethylenediamine (TEMED), zinc nitrate, calcium chloride, and sodium dihydrogen phosphate were all purchased from Sigma (Finland). Tris-HCl buffer solution (pH 8.8 and 6.8), 30% acrylamide/bis, and TBS/Gly/SDS buffer were purchased from Bio-Rad Laboratories, Inc. (Finland). Solutions involved in microRNA-related experiments were prepared with DEPC-treated water (RNase-free) (Sigma, Switzerland). A BCA kit, RIPA, Tris-Triton, protease inhibitor, EDTA, and GelRed were purchased from Thermo Fisher, Finland. DSPE-PEG_2K-RGD, DSPE-PEG-COOH, and RGD were acquired from Xi’an Ruixi Biological Technology Co. Ltd. IR780 and curcumin were purchased from Macklin Inc. (China). Anti-mouse PTEN antibody, (Catalog no. ab267787), Anti-Rabbit GAPDH, (Catalog no. ab210113), Anti-mouse Anti-Cytochrome C, (Catalog no. ab13575), Anti-mouse Anti-beta Actin antibody [mAbcam 8226], Anti-rabbit Anti-pro Caspase-3, (Catalog no. ab32150), Anti-rabbit Anti-Cleaved Caspase-3 (Catalog no. ab32042) were provided from Abcam. Anti-mouse HSP70 Antibody, (Catalog no. #4872) is purchased from Cell Signaling Technology. PMCA ATPase Monoclonal Antibody (Catalog # MA3-914) was provided from ThermoFisher.