GSM7315574: MUT_H3K27ac; Homo sapiens; OTHER
The CUT&Tag assay was conducted using Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme, TD903). Briefly, 105 C4-2 cells were collected and washed once with 500 μl wash buffer before they were bound to ConA beads for 10 min at 25 °C. Then, cells were incubated with IgG, anti-AR (Abcam; ab108341), anti-FOXA1 (Proteintech; 20411-1-AP) and anti-H3K27ac (Cell Signaling Technology; 92964S) at 4 °C overnight. Anti-rabbit IgG was added and incubated for 1 h at 25 °C the next day. Then cells were washed three times with DIG wash buffer and incubated with 0.04 μM pA/G-Tnp for 1 h at 25 °C. Similarly, cells were washed three times with DIG 300 buffer, resuspended in tag mentation buffer and incubated at 37 °C for 1 h. Tag mentation was stopped by adding proteinase K, buffer LB and DNA extract beads. After incubation at 55 °C for 10 min, cells were plated on a magnet and unbound liquid was removed. Beads were gently rinsed twice with 80% ethanol and DNA was eluted with double-distilled water. Libraries were constructed using 'TD903 Hyperactive Universal CUT&Tag Assay Kit for Illumina' and 'TD202 TruePrep Index Kit V2 for Illumina' from Vazyme Biotech.