Jq1 hy 13030

U2OS cells were treated with
6% 1,6-hexanediol (240117, Sigma) prepared in DMEM for 1 min; JQ1
(HY-13030, MedChemExpress) was added to U2OS cells until its final
concentration reached 1.0 μM, and the cells were incubated for
12 h before immunofluorescence microscopy imaging. The immunofluorescence
microscopy experiments were performed following previously published
procedures.^{14 (link)} Briefly, cells on the coverslip
were fixed in methanol/acetic acid (3:1, v/v) for 10 min, permeabilized
with 0.1% triton-X100/PBS for 15 min, and treated with 50 μg/mL
RNase A (EN0531, Thermo Fisher). After blocking with 2% BSA at room
temperature for 1 h, immunofluorescence microscopy experiments were
conducted using standard methods with BG4 (MABE917, Sigma-Aldrich),
anti-FLAG (14793S, Cell Signaling Technology), and anti-rabbit Alexa
594-conjugated (A11037, Invitrogen) antibodies. Nuclei were stained
with DAPI (D9542, Sigma). Finally, the coverslips were mounted with
ProLong Diamond Antifade Mountant (Invitrogen). Images were recorded
using an LSM880 confocal laser scanning microscope (Carl Zeiss) with
a 100× objective and analyzed with ZEN. The foci number per nucleus
were counted using Find maxima in ImageJ. The graphs were plotted
using GraphPad Prism8.