Lactate dehydrogenase assay kit

LDH was measured using a lactate dehydrogenase assay kit according to the manufacturer's protocol (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). SH-SY5Y cells at 1 × 10⁵ cells/well were cultured in 6-well plates. The groups were prepared as above. The culture medium was collected and the level of extracellular LDH measured. The cells were harvested to measure the level of intracellular LDH activity.

Chlorin e6 was purchased from J&K Scientific Ltd. (Beijing, China). Diclofenac, morpholin-4-yl-acetic acid hydrochloride, 1-(3-dimethylaminopropyl)-3 ethylcarbodiimide hydrochloride (EDC), 4-dimethylaminopyridine (DMAP) and trimethylamine (TEA) were purchased from Aladdin (Shanghai, China). Bilirubin and N,N′-diisopropylcarbodiimide (DIC) were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). mPEG_2k-NH₂ and Fmoc-NH-PEG_2k-NH₂ were purchased from Ponsure Biotechnology (Shanghai, China). Cou-6 was obtained from Sigma–Aldrich (USA). DiD and DCFH-DA were obtained from Meilunbio (Dalian, China). The annexin V-fluoresceine isothiocyanate (FITC) apoptosis detection kit and calcium-AM were obtained from Yeason (Shanghai, China). Lactate detection kit and lactate dehydrogenase assay kit were obtained from Nanjing Jiancheng Bioengineering Institute (Naijing, China). Anti-HIF-1α antibody, Cy3 goat anti-rabbit IgG (H + L) and Alexa fluor 647 goat anti-rabbit IgG (H + L) secondary antibody were purchased from Abcam (Hongkong, China). Anti-LDHA and anti-CD31 antibody were purchased from Servicebio Company (Wuhan, China). Anti-LDHB and anti-c-MYC antibody were purchased from Proteintech (Wuhan, China). Anti-VEGFα and anti-angiopoietin-2 antibody were purchased from Abclonal Company (Wuhan, China).

The MDA and LDH levels of the cells were determined using Malondialdehyde (MDA) assay kit (cat. A003-1-2; Jiancheng) and Lactate dehydrogenase assay kit (cat. A020-1-2; Jiancheng) according to the instructions.