Fig. 5: Btn2 SUMOylation restricts INQ accumulation.
From: Dynamics of DNA damage-induced nuclear inclusions are regulated by SUMOylation of Btn2
a Localization of Btn2 is unaffected by the 6KR mutation. b Fractionation of Btn2-WT and Btn2-6KR cells reveals increased insolubility for Btn2-6KR in the pellet fraction. Shown is a graph measuring relative fold change for pellet fraction Btn2 compared to WT from three independent experiments. c Filter trap assay performed using a cellulose acetate membrane retained more Btn2 in Btn2-6KR cells indicating increased insolubility due to loss of SUMOylation. Relative fold change was calculated between equal amounts (20 µg) of protein loaded from three independent experiments (lower left panel). Equal amounts of whole cell lysates were run on an SDS-PAGE and quantified to control for protein levels (lower right panel). Relative fold change was calculated from five independent experiments. d Loss of Btn2 SUMOylation causes elevated sequestration for multiple INQ proteins. Representative images are shown below. e Fractionation of Rpd3 in both Btn2-WT/Btn2-6KR strains in untreated and MMS treated cells. f Btn2-6KR growth effects in a fes1Δhsp104Δ background as seen from a spot dilution assay. Equal ODs of the indicated strains were serially diluted and spotted on YPD at 25 °C, 30 °C, 34 °C and 37 °C. All error bars represent ±SEM, n = 3 biologically independent replicates, >100 cells each. ****p < 0.0001, **p < 0.002, *p < 0.03, Fisher’s test. Scale bars: 5 µm. Source data are provided as a Source Data file.
