Fig. 1
Detection and quantification of PPIs at a single cell level by PLIC. a A graphical scheme illustrating the main steps of the PLIC protocol. FS fluorescent signal. b A graphical scheme illustrating PLIC analysis of Aire and Sirt1 interaction. c PLIC analysis showing representative images obtained from imaging flow cytometry of the interaction of Aire and Sirt1 in mTEChi and CD45+ populations presented as (left to right): bright field (BF), PLA signal (green), staining of CD80 (PB), EpCAM (APC), CD45 (APC/Cy7), and BF/PLA/EpCAM overlay (merged). d Quantitative analysis of Aire and Sirt1 interaction (data are presented as percentage of positive cells (PLA+) in mTEChi or CD45+ cells isolated from WT mice, compared to probes background. Shown are values from three independent experiments. Each point on the graph (triangle for mTEChi, circle for CD45+, and cross for probes only) represents an averaged value of single independent experiment, calculated from 2 to 4 biological replicates, +s.e.m). e PLIC analysis showing representative images of the interaction of Aire and Sirt1 in mTEChi overlaid with Aire staining (left to right): bright field (BF), Aire-AF488 (5H12), PLA signal (FarRed), BF/Aire/PLA/EpCAM overlay (merged), staining of EpCAM (APC/Cy7), CD80 (PB), and CD45 (PE/Cy7). Scale bar 7 μM
