Research Paper Volume 11, Issue 24 pp 12295—12314

Figure 4. ERK1 repressed YAP1 signaling-related gene expression in breast cancer cells. (A) Western blotting showed that silencing of ERK1 increased YAP1 protein levels in T47D cells. (B) Western blotting showed that silencing of ERK1 increased YAP1 protein levels in MCF7 cells. (C) Western blotting showed that silencing of ERK1 did not change p-LATS1/2, LATS1 and LATS2 protein levels in T47D cells. (D) Western blotting showed that silencing of ERK1 did not change p-LATS1/2, LATS1 and LATS2 protein levels in MCF7 cells. (E) Immunofluorescence showed that ERK1 silencing increased YAP1 protein expression in T47D cells. (F) The CHX chase assay showed that the YAP1 protein stability was not altered upon silencing of ERK1 in T47D cells. (G) The CHX chase assay showed that the YAP1 protein stability was not altered upon silencing of ERK1 in MCF7 cells. (H) RT-qPCR showed that ERK1 silencing elevated YAP1 mRNA levels in T47D cells and MCF7 cells. (I) RT-qPCR showed that ERK1 silencing elevated mRNA levels of YAP1 downstream genes (BIRC5, GLS, SGK1) in T47D cells. (J) RT-qPCR showed that ERK1 silencing elevated mRNA levels of YAP1 downstream genes (BIRC5, GLS, SGK1) in MCF7 cells. (K) Knockdown of ERK1 increased GTIIC reporter activity in T47D cells and MCF7 cells. (L) Lentivirus mediated knockdown of ERK1 increased YAP1 protein expression in T47D cells. (M) Lentivirus mediated knockdown of ERK1 increased YAP1 protein expression in MCF7 cells. (N) Lentivirus mediated knockdown of ERK1 increased YAP1 mRNA expression in T47D cells. (O) Lentivirus mediated knockdown of ERK1 increased YAP1 mRNA expression in MCF7 cells. *, p<0.05; **, p<0.01; ***, p<0.001.