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. 2016 Jan 21;18(1):10–24. doi: 10.1016/j.neo.2015.11.010

Figure 3.

Figure 3

AMOTL1 expression is negatively regulated by Merlin. (A) The expression levels of AMOTL1 and Merlin proteins in a set of 16 breast cancer cell lines display a clear opposite pattern. (B) The levels of Merlin and AMOTL1 quantified by Western blot are negatively correlated (Spearman's test). (C) The level of AMOTL1 increases when Merlin protein expression is inhibited in BC52 and MDA-MB-468 using shRNA (sh Merlin #1 and #2 and control shRNA, sh ctrl). (D) The inhibition of Merlin expression in BC52 cells (sh Merlin #1) leads to the upregulation of AMOTL1 mRNA levels measured by real-time PCR (mean ± SD) (Mann-Whitney test). (E) The transcriptional activity of YAP/TAZ was measured in BC52 cells expressing shRNA control or shRNA against Merlin using the YAP/TAD-responsive reporter 8xGTIIC-lux (mean ± SD) (Mann-Whitney test). (F) Real-time PCR analysis of the AMOTL1 transcript levels upon siRNA-mediated YAP inhibition in BC52 cells where NF2 is downregulated by specific shRNA (sh Merlin #1 and #2, control shRNA: sh ctrl) (left panel) (Mann-Whitney test). Immunoblot analysis of AMOTL1 and YAP protein level upon siRNA-mediated YAP inhibition in BC52 cells stably expressing shRNA control (sh ctrl) of NF2 (sh Merlin #1 and #2) (right panel). (G) Correlation between AMOTL1 transcript levels and YAP protein levels (measured by RPPA) in a set of 126 human breast tumors (Spearman test).