Figure 1

Gastric cancer-derived MSCs recruit, protect, and activate neutrophils. Neutrophils were incubated in the presence or absence of GC-MSCs conditioned medium for 18 h, the viability of neutrophils was determined by using MTT assay (a); The apoptosis of neutrophils was quantified by using an Annexin V/PI apoptosis detection kit (b); Total protein was extracted from neutrophils and the levels of Bax and Bcl-2 were detected by using western blot. GAPDH was used as loading control (c); Total RNA was extracted from neutrophils and real-time PCR was performed to determine the mRNA levels of Mcl-1 and FAS (d), IL-8, CCL2, TNFα, and OSM (f, g); Neutrophils were seeded on the upper chamber of a transwell system with 3 μm pore inserts. The bottom chambers were loaded with conditioned medium from GC-MSCs. After 3 h of culture, migrated cells were counted by using the Beckman Coulter HMX (e). *P<0.05, **P<0.01