Rhstc1

The human BMMCs from healthy donors were isolated by density gradient centrifugation using lymphocyte separation medium (TBDscience). To analyze the effects of STC1 on monocytic differentiation, BMMCs were cultured with X-VIVO™ 15 Serum-free Hematopoietic Cell Medium (LONZA; 04-418Q) containing 50 ng/mL hSCF (R&D Systems; 255-SC-050) or/and 200 ng/mL rhSTC1 (R&D Systems; 9400-SO-050) in 5% CO₂ and humidified atmosphere at 37 °C for 3 days. To analyze the effect of STC1 on CD62L^hi CD8⁺ T_naïve cells, BMMCs were cultured with X-VIVO™ 15 Serum-free Hematopoietic Cell Medium (LONZA; 04-418Q) containing 200 ng/mL hIL-2 (R&D Systems; 202-IL-050) or/and 200 ng/mL rhSTC1 (R&D Systems; 9400-SO-050) in 5% CO₂ and humidified atmosphere at 37 °C for 3 days.

MHCC-97H was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in DMEM comprising 10% FBS. SNU-398 was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultivated in RPMI 1640 Medium (GIBCO) with 10% FBS. The recombinant human STC1 protein (rhSTC1) and STC1 neutralizing antibody (STC1-Ab) were purchased from R&D Systems. And RO4929097 was purchased from MedChemExpress (MCE).