Supplementary Figure 1: Gating strategy for flow cytometric analyses.

Wildtype HeLa cells (i.e., no viral transduction with Control-MITO or HA-MITO constructs) were used to define flow cytometric gates. Illustrative plots are shown. On the left, the initial gate for single, viable cells (polygon) was set with 89.6% of events included for further analysis; the forward scatter (area) and side scatter (area) axes are linear. On the right, the second gate (i.e., for EGFP-positivity) was set with 99.92% of wildtype HeLa cells excluded and is shaded green. The light blue zone thus represents background EGFP signal. The forward scatter (area) axis is linear and the EGFP (area) axis is logarithmic. For each sample, 100,000 cells were analyzed on a FACSAria IIU SORP sorter (BD Biosciences) using the FACSDiva software (BD Biosciences) and the data was converted into contour plots with outliers using the FlowJo software (FlowJo). For this figure, HeLa cells were cultured in complete DMEM, authenticated by the Duke University DNA Analysis Facility, and tested for mycoplasma contamination.