Extended Data Fig. 7: RSICS measurements of Mandi CIP system.
From: Mandipropamid as a chemical inducer of proximity for in vivo applications
a, Schematic of RSICS in the ARICS framework. A 256×256 pixel image stack of 300–400 frames acquired in 23 spectral channels is decomposed into three three-dimensional image stacks for GFP (G), YFP (Y) and mCherry (Ch), using a spectral filtering algorithm. An arbitrary ROI delimiting a homogeneous region in the cytoplasm is selected and RSICS analysis is applied to each frame of each image stack. b,c CCFs in the three cross-correlation channels obtained from three-color RSICS measurement on COS-7 cells co-expressing EGFP-PYL, YFP-PYRMandi and mCherry-ABI performed 15 min after incubation with 5 µM ABA-AM (b) or 5 µM Mandi (c). d, Binding efficiencies for the Mandi/ABA CIP systems in the presence of 5 µM ABA-AM or 5 µM Mandi. For comparison, the binding efficiencies obtained in the negative (neg.) and positive (pos.) cross-correlation control samples are shown. Small symbols: individual data points corresponding to RSICS measurement in a single cell. Large symbols: means from experiments. Mean±SD across experiments indicated by horizontal lines. Data pooled from two independent experiments with 13 (neg.), 18 (ABA-AM), 19 (Mandi) and 16 (pos.) cells. e, Binding efficiency of PYRMandi vs. mCherry-ABI in the presence of 200 nM ABA-AM measured by two-color RSICS. For comparison, the binding efficiencies obtained in the negative (neg.) and positive (pos.) cross-correlation control samples measured under identical conditions are shown. Mean±SD across experiments indicated by horizontal lines. Data pooled from 2 (neg, pos) or 3 (ABA-AM) independent experiments with 28 (AB-AM), 10 (neg) and 20 (pos) cells.
