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. 2023 Feb 13;24:25. doi: 10.1186/s13059-023-02867-3

Fig. 6.

Fig. 6.

H3.3 promotes binding of de novo expressed TFs. A–C Immunoblot of whole cell lysates from H3f3afl/fl;H3f3bfl/fl;GtROSA26CRE-ERT2 ESCs treated with either EtOH or 2 μM 4-Hydroxytamoxifen for 48 h followed by treatment with either DMSO or 0.5 μg/ml doxycycline for 48 h showing expression levels of H3.3 and Flag-tagged A BHLHA15, B HAND2, or C TWIST2. *Non-specific band ~30 kDa observed in all lysates. Total histone H3 or β-tubulin are used as loading controls. D–F CUT&Tag average profiles of D BHLHA15-Flag, E HAND2-Flag, and F TWIST2-Flag at promoter-proximal regions enriched in WT ESCs in WT and H3.3 KO ESCs. Three kilobases around the center of promoters are displayed for each analysis, and the number of regions profiled is indicated