Mnng hos

The human osteosarcoma cell lines SJSA-1 and 143B were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). The U2-OS and MNNG/HOS cells were purchased from the Chinese Academy of Sciences Cell Bank (Shanghai, China). Osteosarcoma cells were grown in DMEM (Invitrogen, Waltham, MA, USA) supplemented with 10% FBS (ExCell Bio, Shanghai, China) and 1% penicillin−streptomycin (HyClone, Logan, UT, USA). Human brain vascular pericytes (HBVPs) were purchased from ScienCell Research Laboratories (San Diego, CA, USA) and cultured in Pericyte Medium containing 2% FBS, 1% pericyte growth supplement, and 1% penicillin–streptomycin. These cells were incubated in a humidified environment containing 5% CO₂ at 37 °C. Human immortalized osteoblast hFOB 1.19 cells (Procell Life Science&Technology Co., Ltd.) were cultured in DMEM/F12 (Invitrogen) containing 0.3 mg/mL G418 (TargetMol, Boston, MA, USA) at 34 °C. The cells were routinely confirmed to be negative for mycoplasma.

The human osteosarcoma cell lines MG63, MNNG-HOS and Saos-2 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The human osteosarcoma cell line U-2OS was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). These cell lines were cultured following the ATCC protocols. Standardized culture conditions had been described previously [18 (link)].
The antibodies used were S1PR3 (ab126622; Abcam, Cambridge, UK), YAP (ab52771; Abcam, Cambridge, UK), p-YAP (ab76252; Abcam, Cambridge, UK), c-Myc (ab32072; Abcam), Ki67 (GB13030; Servicebio, Wuhan, China), β-actin (M1210-2; Hua'an Biology, Chuzhou, China), GAPDH (bsm-33033M; Bioss, Beijing, China), anti-rabbit IgG light chain (ab99697, Abcam), anti-rabbit IgG heavy chain (ab99702, Abcam), and anti-mouse IgG light chain (A25012, Abbkine, CA, USA).