(A) Meta-gene analysis of CUT&Tag for WT or ΔARM HA-tagged KLF4 or SOX2, centered on called WT peaks in mESCs
(B) Example tracks of CUT&Tag (spike-in normalized) at specific genomic loci.
(C) Diagram of KLF4 and its cross-correlation to the Tat ARM (magenta), predicted disorder (black line), DNA-binding domain (grey boxes) and predicted disordered domain (cyan).
(D) Side and top views of the crystal structure of KLF4 with DNA (PDB: 6VTX) or AlphaFold predicted structure (ID: O43474)
(E) Experimental scheme for TF gene activation assays. KLF4 ZFs are replaced either by GAL4 or TetR DBD. The effect of KLF4-ARM mutation or replacement of KLF4-ARM with Tat-ARM on gene activation is tested by UAS or TetO containing reporter system.
(F) Normalized luminescence of gene activation assays, normalized to the “No TF” condition (error bars depict s.d., GAL4: p<0.0001 for all pairwise comparisons except WT vs. Tat-ARM, p=0.3363; TetR: NoTF vs. WT, p<0.0001, NoTF vs. R/K>A, p=0.5668, NoTF vs. Tat-ARM, p=0.0002, WT vs. R/K>A, p=0.0003, WT vs. Tat-ARM, p=0.7126, Tat-ARM vs. R/K>A, p=0.0008, one-way ANOVA)