GSM8423521: H226R_7883_KDFOSL1_IgG_1_; Homo sapiens; OTHER
H226/ H226-7883R cells were trypsinized into single-cell suspension with 0.05% Trypsin, EDTA 0.5 mM.  500k cells were used for each reaction. We used the CUTANA ChIC/CUT&RUN Kit from Epicypher (#14- 1048) following manufacturer's instructions. ConA Beads were briefly resuspended and 11 μl of beads per  reaction was transferred to a 1.5 ml tube. Beads were resuspended in 100 μl bead activation buffer per  reaction and washed 2✕. Then resuspended in 11 μl buffer per reaction and aliquoted 10 μl into 8-strip PCR  tubes, and kept on ice. Harvested cells were spun down at 600 × g for 3 min at room temperature. Cells  were then resuspended in 100 μl/reaction of wash buffer and aliquoted into 8 strip tubes with the 10 μl of  activated ConA beads. After 10 mins incubation, 50 μl of cold antibody buffer was added. YAP antibody  (CST Cat. #14074) was used at 1:50 dilution, TEAD1 antibody (BD Cat. #610923) or FOSL1 antibody  (ThermoFisher Cat. #PA5-66880) was added at 0.5 μg per reaction. Tubes were incubated on nutator  overnight at 4 °C. The next day, tubes were collected and washed 2✕ with 200 μl cell permeabilization  buffer. Beads were resuspended in 50 μl cell permeabilization buffer and 2.5 μl pAG-MNase was added.  After 10 min incubation at room temperature, beads were washed with another 200 μl cell permeabilization  buffer 2✕. Finally beads were resuspended in 50μl cell permeabilization buffer. 1 μl of 100 mM CaCl2 was  added to activate the enzyme. Tubes were then incubated for 2 h on nutator at 4 °C. Finally, the reaction  was stopped with 33 μl of stop master mix. DNA was then purified and quantified using a Qubit fluorometer.  For library preparation, we used the CUTANA CUT&RUN Library Prep Kit from Epicypher (#14-1001). End  Repair step, Adapter ligation and U-excision were performed following manufacturer's instructions. We  used 1✕ SPRIselect beads for DNA cleanup. After indexing PCR, we then used 1✕ SPRIselect beads for  DNA cleanup prior to sequencing. Analysis of the library fragment size was done using Tapestation.  10 million Paired End Reads (PE) per sample.