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

Figure 4.

Figure 4

Both JNK and ERK pathways contribute to FRA-1 expression through MLK3 signaling. (a) MCF7 cells were transiently transfected with a wild-type MLK3 (pRK-MLK3) or a kinase dead MLK3 variant (pRK-MLK3-K144M) for 24 h, (b) SUM-159 and (c) 4T1 cells were treated with vehicle, 400 nm CEP-1347 (MLK inhibitor), 15 μm SP600125 (JNK inhibitor), 10 μm U0126 (MEK/ERK inhibitor) or 10 μm SB203580 (P38 inhibitor) for 24 h. Cellular lysates were subjected to immunoblotting with indicated antibodies. Western blot quantification of the indicated protein normalized to actin is expressed as mean±s.d. from at least three independent experiments. (d) The mRNAs from 4T1 cells treated with vehicle, 400 nm CEP-1347, 15 μm SP600125 and 10 μm U0126 for 24 h were subjected to qRT–PCR analysis with FRA-1 primers. Relative mRNA expression is displayed as mean±s.d. from at least three independent experiments performed in triplicate; NS, not statistically significant; **P<0.01.