Extended Data Fig. 10: Examples of bifunctional domain sequences at three different promoters.

a, Tiling plot for LEUTX (n = 2, dots are the mean, bars the range). b, Deletion scan across one of LEUTX’s bifunctional tiles (n = 2, dots are the mean, bars the range). Deletions were binned by their statistical significance into those that decreased activity (gray lines) compared to the WT tile and those that did not (one-tailed z-test). The necessary sequence for another gene family member, ARGFX, is highlighted in teal. c, Bifunctional domain necessary region location categories. Overlapping regions were defined as any tile that contained a deletion that was both necessary (below activity threshold) for activation and necessary for repression. d, Citrine distributions of ARGFX-161:240 recruited to minCMV (n = 2, left), and recruited to pEF (n = 2, right). e, Citrine distributions of bifunctional tiles identified from minCMV and pEF CRTF tiling screens recruited to PGK promoter (n = 2). Asterisks denote p-values < 0.05 for the percentage of cells on (right) and off (left) in the dox population (one-sided Welch’s t-test, unequal variance). ARGFX-191:270 off p = 0.0003, on p = 0.02; FOXO1-561:640 off p = 0.017, on p = 2.44e-5; NANOG 191:270 off p = 2.12e-5, on p = 0.0002; NANOG 225:304 off p = 0.202, on p = 0.0004; KLF7 1:80 off p = 0.99, on p = 0.0005. f, Comparison between set of proteins screened in Alerasool et al., 2022’s ORFeome and this study.