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. 2018 Aug 7;6:86. doi: 10.3389/fcell.2018.00086

Figure 4.

Figure 4

The interaction between CADM1 and CADM4 is involved in the cell spreading of ATN1 cells. (A) The trans-interaction between CADM1 and CADM4 in the extracellular region was examined by immunoprecipitation assay. E-cadherin and CADM3 were used as the negative and positive controls, respectively. (B) Knockdown of CADM1 in ATN1 cells by two independent shRNAs, shCADM1-5, and shCADM1-8. (C) ATN1 cells with control shRNA or shCADM1 were incubated on coverslips coated with control IgG or CADM4-Fc. The area of 100 cells was measured in an assay and the average of three independent experiments is shown. *p < 0.05 by t-test. (D) Representative images of the spread morphology of ATN1 cells with each shRNA incubated on IgG- or CADM4-Fc-coated glasses. The cells were visualized by staining the actin cytoskeleton with Alexa Fluor 488-labeled phalloidin. The nuclei were stained with DAPI. Magnification, × 200. Scale bar, 20 μm. (E) ATN1/shControl cells were incubated on IgG or CADM4-Fc for 60 min in the presence of control chicken IgY (10 μg/mL) or anti-CADM1 antibody, 9D2 (10 μg/mL). The average cell area of three independent experiments is shown. *p < 0.05 by t-test.