Rabbit igg

All chemicals, unless otherwise noted, were obtained from ThermoFisher or Merck. Enzymes were obtained from New England Biolabs. The following drugs/dyes were used for this work: IFNγ (final concentration of 50 ng/ml, Merck), dTAG13 (final concentration of 100 nM, Merck), Vybrant DyeCycle Ruby Stain (final concentration of 5 μM, ThermoFisher) and Tofacitinib citrate also known as CP‐690550 (JAK inhibitor, concentration of 10 μM, Merck). The following antibodies were used for this work: GBP5 (Cell Signaling Technology, 67798; Abcam, ab96119), STAT1 (Cell Signaling Technology, 9172), Phospho‐STAT1 (Ser727) (Cell Signaling Technology, 9177), Phospho‐STAT1 (Tyr701) (58D6) (Cell Signaling Technology, 9167), STAT2 (Santa Cruz Biotechnology, sc‐1668), STAT3 (Santa Cruz Biotechnology, sc‐8019), STAT5B (Santa Cruz Biotechnology, sc‐1656), IRF1 (Cell Signaling Technology, 8478), IRF9 (ThermoFisher Scientific, 702322), a‐TUB (Merck, T9026), anti‐rabbit (fluorophore‐conjugated) (LI‐COR, 926‐32211), anti‐mouse (fluorophore‐conjugated) (Rockland, 610‐744‐124), Rabbit IgG (Epicypher, 13‐0042k).

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)).