Fig. 4

Ada1 is a key factor in Ubp8 cellular localization. a Localization of Ubp8 tagged with GFP in WT, ada1Δ, spt20Δ and ada1Δspt20Δ cells was monitored using fluorescence microscopy. Nuclei were stained with DAPI (DNA). Images were cropped from full sized frames of ~ 250–500 cells and scale bars are set to 5.0 µm. b Boxplot indicating GFP nuclear intensity for Ubp8-GFP WT, Ubp8-GFPada1Δ, Ubp8-GFPspt20Δ and Ubp8-GFPada1Δspt20Δ strains obtained from more than 100 cells in each case. c Localization of the Sus1 protein tagged with GFP in WT and spt7Δ was monitored using fluorescence microscopy. Nuclei were stained with DAPI (DNA). d Upper panel: levels of Sus1 protein tagged with GFP in WT and spt7Δ mutant cells were analyzed by western blotting of whole-cell extracts using an anti-α-GFP antibody. Levels of Pgk1 protein were monitored as the loading control. Lower panel: serial dilutions of WT and mutant spt7Δ cells, expressing or not expressing Sus1-GFP-tagged proteins as indicated, were grown on YPD plates. Plates were incubated for 48 h at 30 °C