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. 2017 Jun 12;6(6):e345. doi: 10.1038/oncsis.2017.44

Figure 6.

Figure 6

MLK3 regulates cancer cell invasion and transendothelial migration. (a–c) MCF7iMLK3 cells treated with vehicle or with 25 nm AP21967 plus either 50 nm control or FRA-1 siRNA, as indicated, and (d) parental 4T1 cells treated with vehicle or 400 nm CEP-1347 and MLK3-KO 4T1 clones (KO-1, KO-2 and KO-3) were subjected to an in vitro Matrigel transwell invasion assay for 24 h. Relative cell invasion, with control set at 100%, is expressed as mean±s.d. from three independent experiments. (e) A representative image from three independent experiments of gelatin zymography of conditioned medium from parental 4T1 cells treated with vehicle or 400 nm CEP-1347 and MLK3-KO 4T1 clones (KO-1, KO-2 and KO-3). (f and g) Parental 4T1 and 4T1KO-1 were subjected to transendothelial migration toward 10% FBS. Endothelial permeability was assessed using 5(6)-Carboxyfluorescein (6-FAM)-conjugated albumin, and transendothelial migration was assessed using bioluminescence imaging as described in the 'Materials and Methods' section. Relative 6-FAM conjugated albumin concentration and bioluminescence activity are expressed mean±s.d. from three independent experiments performed in triplicate. **P<0.01. *P<0.1. NS, not significant.