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. 2022 Mar 2;50(6):3276–3291. doi: 10.1093/nar/gkac135

Figure 2.

Figure 2.

Chromatin-bound Cse4 contributes to SDL. (A) Reduced dosage of histone H4 (hhf1Δ or hhf2Δ) suppresses the cdc48-3 GAL-CSE4 SDL. The indicated strains containing either vector or GAL-CSE4 were spotted in five-fold serial dilutions on glucose (2%)- or galactose (2%)-containing synthetic medium selective for the plasmid. The plates were incubated at 25°C for 4 days. (B) Overexpression of cse4 K215/216R mutant exhibits reduced SDL compared to GAL-CSE4 in cdc48-3 strain. Growth assay was conducted as described in A. The plates were incubated at 25°C for 5 days. (C) Chromatin-bound endogenous Cse4 is enriched in cdc48-3 and npl4-1 strains. Whole cell extracts (WCEs) prepared from equal numbers of logarithmically growing cells in YPD were fractionated into soluble and chromatin fractions. Cse4 levels in each fraction were monitored by Western blot analysis with anti-HA antibody. Pgk1 and histone H2B were used as markers for soluble and chromatin fractions, respectively.