ChIP assay was done as previously described (Jacob et al, 2011). Briefly, fibroblasts were cross‐linked in 1% formaldehyde for 10 min at RT followed by glycine quenching. Cells were then washed and lysed, and the chromatin was sonicated into 200‐ to 1,000‐bp fragments, diluted, and pre‐cleared using protein A beads. Anti‐Rel‐A‐ (Santa Cruz) or isotype‐matched IgG (control) antibodies were incubated with the chromatin samples overnight. Protein A beads were added for 2 additional hours, and the beads were then washed and the cross‐linked reversed overnight at 65°C. The DNA was purified and subjected to quantitative PCR analysis using the indicated primers. The dissociation curves following amplification showed that all primer pairs generated single products. The amount of PCR product amplified was calculated relative to the input.
Primers:

hTNFA promoter: For: CCTCCAGATGAGCTCATGGGTT, Rev: GGGTGTGCCAACAACTGCCTTT.

hIL1B promoter: For: TGTCTTCCACTTTGTCCCACA, Rev: CGTTGTGCAGTTGATGTCCA.

hp21 promoter: Validated primer set was purchased from Millipore.

hIKB promoter: Validated primer set were purchased from Millipore.

Falik‐Zaccai T.C., Barsheshet Y., Mandel H., Segev M., Lorber A., Gelberg S., Kalfon L., Ben Haroush S., Shalata A., Gelernter‐Yaniv L., Chaim S., Raviv Shay D., Khayat M., Werbner M., Levi I., Shoval Y., Tal G., Shalev S., Reuveni E., Avitan‐Hersh E., Vlodavsky E., Appl‐Sarid L., Goldsher D., Bergman R., Segal Z., Bitterman‐Deutsch O, & Avni O. (2017). Sequence variation in PPP1R13L results in a novel form of cardio‐cutaneous syndrome. EMBO Molecular Medicine, 9(3), 319-336.
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