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. 2024 Jan 5;223(3):e202306013. doi: 10.1083/jcb.202306013

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© 2024 Endo et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure S1. — Ubiquitin accumulation in USP8-depleted cells. (A) HeLa cells transfected with the indicated siRNAs were subjected to subcellular fractionation. Each fraction (cytoplasm, membrane, nucleoplasm, chromatin, and cytoskeleton) was immunoblotted with the indicated antibodies. (B) HeLa cells transfected with the indicated siRNAs were immunostained with the indicated antibodies and DAPI. Scale bar, 20 μm. (C) HeLa cells stably expressing siRNA-resistant Myc-USP8 WT and C786A were transfected with the indicated siRNAs. Cells were immunostained with the indicated antibodies and DAPI. Scale bar, 20 μm. (D) HeLa cells transfected with the indicated siRNAs were immunostained with the indicated antibodies and DAPI. Scale bar, 20 μm. (E) Total cell lysates from parental HeLa cells and HeLa cells stably expressing FLAG-Ub were immunoblotted with the indicated antibodies. (F) HeLa cells stably expressing FLAG-Ub were transfected with the indicated siRNAs. Cells were immunostained with the indicated antibodies and DAPI. Scale bar, 50 μm. Source data are available for this figure: SourceData FS1.