Knockdown of YAP inhibits autophagic flux in breast cancer cells.

<p><b>(A)</b> MCF7-shCtrl and shYAP cells were cultured in normal medium (ND 0 h or no treatment) or maintained in EBSS (ND) +vehicle control (VC) or EBSS +CQ or Bafilomycin A1 (BafA1) for 2,4, and 6 h. Total proteins were subjected to Western blotting for the indicated antibodies. Representative results are shown. <b>(B)</b> Quantified relative LC3-II flux according to the Western blot results of (A) is expressed as the normalized LC3-II ratio calculated by the levels of LC3-II in the presence of CQ divided by the levels of LC3-II in absence of CQ. Data are derived from three independent experiments and shown as the means±SEM. *<i>P</i><0.05. <b>(C)</b> Quantitative relative p62 levels according to results of (A) are expressed as p62 levels after different treatments divided by that of no treatment. Data are derived from three independent experiments and shown as the means±SEM. *<i>P</i><0.05. <b>(D)</b> MCF7-shCtrl, shYAP and YAP-rescue-overexpression cells were cultured in normal medium (ND 0 h or no treatment) or maintained in EBSS (ND)+Vehicle Control (VC) or EBSS +CQ for 6 h. Total proteins were assayed by Western blotting. Representative results are shown. <b>(E)</b> Quantitative relative LC3-II flux according to Western blot results of (D) was assayed as previous. Data are derived from three independent experiments and shown as the means±SEM. *<i>P</i><0.05. <b>(F)</b> Quantified relative p62 levels according to the results of (D) were assayed as previous. Data are derived from three independent experiments and shown as the means±SEM. *<i>P</i><0.05. <b>(G)</b> MDA-MB-231-shCtrl and shYAP cells were cultured for 4 h in normal medium (no treatment), EBSS+VC (ND) or EBSS+BafA1. Total proteins were assayed by Western blotting with indicated antibodies. <b>(H)</b> MDA-MB231-Control and YAP overexpression cells were cultured for 6 h in normal medium (no treatment), EBSS+VC (ND) or EBSS+BafA1. Total proteins were assayed by Western blotting.