CUT&RUN of NFKB1 and JUN in TNF treated RPTEC cells

Analysis of snATAC-seq data suggests increased motif of NFKB and AP-1 motifs of renal epithelia in kidney injury. To confirm binding sites we performed CUT&RUN of JUN and NFKB1 transcription factors in TNF-alpha treated cultured primary human renal proximal tubule cells. Primary human renal proximal tubule cells were cultured and treated with 20ng/ml of TNF-alpha 24hours before harvest. Cells were detached and nuclei isolated. The CUT&RUN assay was then performed using the ChIC/CUT&RUN kit (EpiCypher). Positive reactions were performed using JUN and NFKB1 antibodies. Background signal was assessed using an IgG antibody.