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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: Tumor cell-derived SPON2 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by activating PYK2 in CRC

Fig. 3

SPON2 promotes TAM transendothelial migration. a GSEA identified the leukocyte transendothelial migration signature as the top activated pathway in SPON2-high CRC patients (TCGA COAD). b Representative immunofluorescence for ZO-1 in HUVECs cocultured with THP-1 and treated with conditioned media (CM) from SW480/Vector and SW480/SPON2. Scale bar, 50 μm. c Immunofluorescence of F-actin (green) in THP-1 cells pretreated with conditioned medium (CM) from stable cell lines (SW480/Vector and SW480/SPON2). Nuclei were counterstained with DAPI (blue). Scale bar, 20 μm. d Migration of THP-1 s toward conditioned medium from stable cell lines (SW480/Vector, SW480/SPON2, SW620/ Scramble, SW620/shSPON2#1 and SW620/shSPON2#2). e THP-1 s were stained with DIL (red) before pretreatment with conditioned medium as indicated and were cocultured with HUVECs labeled with pCDH-GFP (green) for 2 h. Cells were observed under a fluorescence microscope and counted from 5 random fields. Scale bar, 100 μm. f Transendothelial migration of THP-1 s toward conditioned medium from stable cell lines. Scale bar, 100 μm. Student’s t test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

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