Fig. 3: HSF2 interacts with CBP and EP300 in normal conditions.

a Schematic representation of CBP protein domains. The ability of CBP to bind a very large number of proteins is mediated by several conserved protein binding domains, including the nuclear receptor interaction domain (RID), the cysteine/histidine-rich region 1 (CH1), the KIX domain, the bromodomain (BD), the CH2 containing a PHD and a RING domain, the HAT, the CH3, the steroid receptor co-activator-1 interaction domain (SID) and the glutamine- and proline-rich domain (QP)³³. b Representative kinetics of recombinant HSF2 binding to His-tagged CBP domains or HSP70 (positive control) by biolayer interferometry (n = 3). c Schematic representation of the principle of the fluorescent-3-hybrid (F3H) assay. Genomic integration of a LacO array allows the focal recruitment in the nucleus of a LacI fused to the GFP binder, which in turn recruits the GFP-tagged probe (HSF2-YFP) and its potential interactants (CBP/EP300), being either endogenous or brought by overexpression. d, e Representative confocal sections of BHK cells carrying a stably integrated Lac-operator array, transfected with LacI-GFP binder, HSF2-YFP, and CBP-HA (d) or EP300-HA (e) showing the interaction between HSF2-YFP (green) and exogenous CBP-HA, endogenous CBP (d) or with exogenous EP300-HA (e) (red). White arrows, co-localization of HSF2 and CBP or EP300 at the LacO array. Negative controls are shown in Supplementary Fig. 3d–g. Scale bar: 10 μm. Graphs represent the combined signal intensity of the two fluorescence signals at the LacO array. Quantification: percentage of cells showing co-recruitment of YFP-HSF2 and EP300-HA, CBP-HA or endogenous CBP at the Lac0 array (n = 3 or 4, average of 100 counted cells per experiment). Source data are provided as a Source Data file.