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. 2022 Apr 18;19(1):152–162. doi: 10.1080/15548627.2022.2063004

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© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — YAP1 and IL6ST are degraded via the lysosomal pathway. (A,B) Treatment of Hep3B cells for 6 h with the lysosomal inhibitor(s) NH₄Cl (20 mM) and leupeptin (50 μM) (A) or with a combination of the two for the indicated time (B) leads to the accumulation of YAP1 and IL6ST in Hep3B cells. SQSTM1 is shown as a positive control. The bottom panel show the fold change of YAP1 and IL6ST over their expression in untreated cells, set as 1. (C,D) Treatment of Hep3B cells for 6 h with the indicated concentration of the proteasome inhibitor MG132 (C) or for the indicated time with 5 μM MG132 (D) has moderate to no effects on YAP1 and IL6ST levels. The bottom panel show the fold change of YAP1 and IL6ST over their expression in cells treated with DMSO, set as 1. (E,F) Silencing of either ATG5 or ATG7 does not cause the accumulation of YAP1 and IL6ST in Hep3B cells. SQSTM1 is shown as a positive control. The bottom panels show the fold change of YAP1 and IL6ST over their expression in cells transfected with siScr, set as 1. (G) Treatment of Hep3B cells for 24 h with the indicated concentrations of the macroautophagy-inducing drug rapamycin fails to reduce the levels of YAP1 and IL6ST in Hep3B cells. SQSTM1 is shown as a positive control. The bottom panel shows the fold change of YAP1 and IL6ST over their expression in cells treated with DMSO, set as 1. In all panels, the intensity of the bands of interest was normalized to TUBA prior to fold-change calculations. Data are plotted as the mean ± SEM of n = 3 biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001 vs control.