Rhodamin 123

Serum-starved hCorECs and hRCECs were exposed to three kinds of PEG-GO nanosheets (50 μg/mL) for 24 h, while untreated cells were used as a control. The MMP and ROS detection were performed as described previously [22 (link),26 (link)]. The cells were washed with PBS three times and then incubated with 1 μM Rhodamin 123 (C2007, Beyotime, Shanghai, China) and 10 mM H2DCF-DA (20, 70-dichlorofluorescin diacetate) (D6883, Sigma, St. Louis, MO, USA) for 30 min to detect MMP and ROS, respectively. After incubation, cells were washed twice with PBS. The fluorescence intensity was then determined by FACS Calibur Flow Cytometer (BD Bioscience, San Jose, CA, USA) and analyzed using FlowJo software (Ashland, OR, USA). Three independent experiments were conducted.

Arsenic trioxide (Sigma, USA) was stocked at 2 mM and stored at 4°C. Nutlin-3 (Cayman, USA) was dissolved in DMSO at 40 mM and stored at -20°C. R (+) Verapamil (Sigma, USA) was stocked at 1 mM and stored at 4°C. Rhodamin 123, JC-1 and DAPI were purchased from Beyotime (Shanghai, China). RPMI1640, DMEM and fetal bovine serum were also purchased from Invitrogen.

Oridonin (≥98%, HPLC) is obtained from Mingwang biotechology (China). Fetal calf serum (FCS), penicillin/streptomycin, dulbecco’s modified eagle medium (DMEM), and trypsin kit are obtained from Gibco (USA). Paraformaldehyde is purchased from Sigma (USA). 3-(4, 5)-dimethylthiazo(-z-y1)-3,5-diphenyt- etrazoliumromide (MTT), N-acetyl-L-cysteine (NAC), Annexin V-FITC/PI (Annexin V-Fluorescein Isothiocyanate/Propidium Iodide) apoptosis detection kit, DCFH-DA (2',7'-dichlorodihydrofluorescein diacetate) reactive oxygen species assay kit, rhodamin 123, actin-tracker green (phalloidin- Fluorescein Isothiocyanate), 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) and cell cycle analysis kits (Propidium Iodide) are purchased from Beyotime Institute of Biotechnology, China.