Supplementary Figure 5: Identification of binding motifs for TF heterodimers using SMiLE-seq.
From: SMiLE-seq identifies binding motifs of single and dimeric transcription factors
a. Schematic representation of the experimental setup. Step 1. Biotinylated anti-eGFP antibody is immobilized under the button of the SMiLE-seq device. Step 2. Dimerizing transcription factor (TF1) fused to an eGFP tag, dimer partner (TF2) tagged with mCherry and Cy5-labeled DNA baits are introduced into the chip. Step 3. Antibody-immobilized complexes consisting of TF1, TF2, and DNA are trapped under the flexible PDMS membrane; dimer formation is confirmed by fluorescent read-out. Step 4. Unbound molecules as well as molecular complexes are washed away. b. TOMTOM3 comparison of JASPAR and SMiLE-seq binding motifs for mouse PPARγ:RXRα and human ARNTL:CLOCK heterodimers.
3. Gupta, S., Stamatoyannopoulos, J. A., Bailey, T. L. & Noble, W. Quantifying similarity between motifs. Genome Biol. 8, R24 (2007).
