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. 2021 Jul 26;22(15):7943. doi: 10.3390/ijms22157943

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Deletion of DSK2s results in the accumulation of ubiquitinated proteins. (a) Schematic representation of ΔΔdsk2adsk2b construction. The dsk2b gene locus in Δdsk2a::CAT strain was replaced by DHFR cassette. (b) Diagnostic PCR of ΔΔdsk2adsk2b construction. PCR1 and PCR2 were used to identify the homologous integration. PCR3 was used to amplify the fragment of dsk2b. (c,d) Western blotting determined the ubiquitinated proteins in ΔΔdsk2adsk2b and Δku80. Ubiquitinated proteins were detected by rabbit anti-ubiquitin (linkage-specific K48) monoclonal antibody (upper panel). Actin served as a loading control (bottom panel). Graphs represent densitometric relative to actin. (mean ± SD from three biologically independent experiments). ** p < 0.01.