Extended Data Figure 6. Secondary motifs promote inclusion in a splicing reporter in an RBFOX1-dependent manner at the protein level.

a. Six secondary motifs approximate the exon inclusion of one primary motif in an Rbfox1-dependent manner at the protein level, replicate 2. RG6 plasmids containing one primary motif or six secondary motifs were co-transfected in HEK293T cells with fluorescently labelled Rbfox1 and monitored by flow cytometry for the inclusion isoform (GFP), exclusion isoform (dsRED), and Rbfox1 (Cerulean) expression at the single-cell level. Controls including a scrambled motif co-transfected with Rbfox1 (light grey) and scrambled and intact motifs without Rbfox1 (grey) are also shown. Bins detailed in Supplementary Table 5. b. The slope of linear fit of two flow cytometry replicates were null-subtracted and normalized to their permuted controls. Error bars represent standard error of the mean (SEM).