The SIR complex is susceptible to lipid alterations at the NE. (A) Wild-type (W303) cells or the indicated SIR complex mutants were serial diluted onto synthetic solid media containing 25 μM edelfosine or vehicle and incubated at 30°C for 2 d. (B) ChIP-qPCR of Sir4^MYC after 60 min in edelfosine or the vehicle. The fold enrichment at three native sub-telomeres (Tel01L, Tel06R, and Tel15L) is shown, normalized to a late replicating region on Chromosome V (469104–469177). Bars represent mean ± SD for four independent experiments while circles represent individual experiments. * indicates P value <0.05; ** indicates P value <0.01 as determined by unpaired t tests with Holm-Šídák correction for multiple comparisons. (C) Representative images of cells expressing Sir4^GFP and the ER marker ^DsRedHDEL (both endogenously tagged) visualized by live fluorescence microscopy after 90 min with edelfosine or vehicle. Scale bar represents 2 µm. The distribution of values from the quantification of the number of Sir4 foci per cell is shown beside the microscopy images. Values cumulative from three independent experiments (n = 180 cells per treatment) are shown as small circles, with the means of each experiment shown as large circles. Bars represent the mean with 95% confidence interval from the pooled values. * indicates a P value <0.05 as determined by two-tailed nested t test (N = 3). For all panels: Ctrl = vehicle; EDLF = edelfosine.