Figure 6

GC-MSCs-primed neutrophils induce the CAF differentiation of normal MSCs. (a) GCN-MSCs were incubation with conditioned media from GC-MSCs-primed neutrophils for 24 or 48 h. Total protein was extracted and the levels of α-SMA and vimentin were detected by using western blot. (b) GCN-MSCs were incubated with conditioned medium from GC-MSCs-primed neutrophils for 48 h. Then the cells were incubated with anti-α-SMA or anti-vimentin primary antibodies followed by fluorescence-conjugated secondary antibodies. Nuclei were visualized by DAPI staining. The images were acquired by using a Nikon eclipse Ti-S microscope. Scale bar= 100 μm. (c)Total RNA was extracted from GCN-MSCs and real-time PCR was performed using human-specific primers for the quantification of α-SMA, FAP, and IL-6. (d) Cell migration assay for SGC-7901 cells stimulated with the conditioned medium from CAF-differentiated MSCs that were induced by GC-MSCs-primed neutrophils. Scale bar=200 μm. **P<0.01