YAP interferes with autophagic flux via enhancing autolysosome degradation.

<p><b>(A)</b> MCF7-shCtrl and shYAP cells were cultured for 6 h in normal medium (no treatment), EBSS (ND), EBSS+3-MA or EBSS+CQ. Total proteins were subjected to Western blotting. Representative results are shown. <b>(B)</b> Quantitative relative levels of LC3-II according to the results of (A) are expressed as LC3-II levels after different treatments (ND, ND+3-MA, ND+CQ) 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>(C)</b> Quantitative relative p62 levels according to the 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 Res cells were treated with rapamycin (Rapa) or Rapa +CQ for 4 h. Total proteins were subjected to western blotting. Increased LC3-II levels were quantified as LC3-II levels after different Treats (Rapa, Rapa+CQ) subtract that of no treat. Quantitative data are derived from three independent experiments and shown as the means±SEM. *<i>P</i><0.05. <b>(E)</b> Lysosomal acidification levels of MCF7-shCtrl, shYAP and YAP Res cells were assayed by flow cytometry after staining with LysoTracker Red. <b>(F)</b> ACP activities of MCF7-shCtrl, shYAP and YAP Res cells were assayed. <b>(G)</b> MCF7-shCtrl-tfLC3 and MCF7-shYAP-tfLC3 cells were starved in EBSS for 2 h. CQ was then added to elevate the lysosomal pH. Images of live cells were taken with a confocal microscope (Scale bar: 10 μm). <b>(H)</b> Percentages of cells with red puncta per field were counted and calculated. Data are shown as the means±SEM. *<i>P</i><0.05, n = 3. <b>(I)</b> Percentages of cells with more than half the puncta yellow per field were counted and calculated. Data are shown as the means±SEM. *<i>P</i><0.05, n = 3.