Ab37264

Bladder tissue microarrays, containing 40 cases of bladder cancer tissues with different stages and grades of cancer and 8 cases of normal tissues, were purchased from US Biomax Inc. (Rockville, MD, USA). Paraffin sections were stained with macroH2A1 (ab-37264, Abcam) and Lin28B (ab-71415, Abcam) antibodies at 4 °C overnight. Then, the sections were incubated with secondary antibodies at room temperature for 1 h, followed by incubation with ABC solution (ABC reagent Elite Kit, Vector Laboratories, Burlingame, CA, USA) at room temperature for 1 h. After staining with diaminobenzidine, images were acquired using a Nikon ECLIPSE 80i microscope (Nikon). A scoring for immunohisochemical staining: negative or marginal (<20% of cells), weak (20–50% of cells), moderate (50–80% of cells) and strong (>80% of cells).

2 × 10⁵ short-term cultured CAFs before or after 30 min of serum stimulation were processed for CUT&RUN^{158 (link)} for macroH2A1 (antibody ab37264, Abcam, lot number GR278020-1, 1 μg per reaction). Cells were permeabilized with 0.0085% digitonin and incubated with antibody. DNA was cleaved with the CUTANA pAG-MNase (Epicypher). Released DNA was processed using a NEBNext Ultra II Library Preparation kit for Illumina (NEB), including a 14-cycle amplification step. Libraries were sequenced and reads were processed as for ATAC–seq.