Screening platform for finding mutants that affect the age-associated decline of MMP. (A) Schematic of the screen. Image of sequencer courtesy of Illumina, Inc. (B) Raw data of the sort showing gating strategy. (C) After 3 (T3) and 24 (T24) hours, samples of the population were imaged on a fluorescent microscope and their bud scars were counted (T3, n = 36; T24, n = 34). (D) Correlation of barcode counts in two independent experiments (“190327” and “190410”). (E) Correlation between deletion mutants whose frequency significantly changed (adjusted P < 0.05) in the high vs low MMP ratio sorts. Error bars show 1 SEM. (F) Correlation between the MMPs portion of the fluorescent reporter, adjusted for mitochondrial mass (Materials and methods), and the MMP-sensitive dye TMRM in WT (orange) and each of the rescreened deletion mutants (blue). Error bars show 1 SEM. Plot shows data from six independent experiments. See Materials and methods for details. In D–F, the dashed line indicates best-fit line from Deming regression (Materials and methods). “r” indicates the Pearson correlation coefficient, and “p” is the P-value of r.