Rabbit anti yy1

Rabbit anti-YY1 (Cell Signaling, Tech), Rabbit anti-KLF4 (Novus Biologicals), anti-rabbit IgG, Isotype control, AlexaFluor 488 Streptavidin (Jackson-lmmunoresearch, West Grove, CA, USA) and Vectashield-DAPI (Vector-laboratories, Burlingame, CA, USA) were used to stain Ramos cell lines transfected with siRNA-YY1. Images were acquired using a Leica TCS SP8x Confocal Microscope (Wetzlar, Germany) and were analyzed with Leica software.

After 48 hours, the transfected HCCs were fixed in 4% formaldehyde, the cells were permeabilized with 0.1% Triton X‐100 and stained using rabbit anti‐YY1(Cell Signaling Technology; diluted 1/500) and mouse anti‐HDAC1 antibody (Cell Signaling Technology; diluted 1/1500), and the cells were measured by a Nikon Eclipse TS100 microscope (Nikon).

CUT&RUN was performed with the CUTANA ChIC/CUT&RUN Kit (Epicypher, 14-1048). A total of 500,000 cells for each condition were fixed with 1% formaldehyde for 1 min, and the protocol for fixed cells was followed as described in the manual. Antibodies used were rabbit IgG (Epicypher, 13-0042), rabbit anti-YY1 (Cell Signaling Technology, 46395S) at 1:50, and rabbit H3K27ac (ActiveMotif, 39034) at 1:50. Libraries were prepared using the CUTANA ChIC/CUT&RUN Library Prep Kit (Epicypher, 14-1001). Samples were run on the Illumina NextSeq 1000 P2 flow cell, with 50-bp paired end reads. Sequenced reads were analyzed using the nf-core/cutandrun pipeline (20 (link), 21 (link)). Peak calling was performed using SEACR (22 (link)).