ChIP Analysis of c-Myb Variants

The HD-11 cell line was transfected with plasmids encoding TY-tagged c-Myb variants (empty vector, 3xTY1-c-Myb, 3xTY-c-Myb D152V) described in [13 (link)]. The ChIP analysis was performed essentially as previously described [53 (link), 54 ], using anti-Ty1 antibodies for IP of c-Myb and anti-H3K27ac antibody (# 39,133, Active Motif) for IP of H3K27ac. The anti-Ty1 monoclonal mouse antibody was produced in our lab from a hybridoma cell line [55 (link)]. Occupancy was determined with qRT-PCR using primers for specific regions of the mim-1 locus (LECT2). Primer sequences are available upon request.

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Fuglerud B.M., Ledsaak M., Rogne M., Eskeland R, & Gabrielsen O.S. (2018). The pioneer factor activity of c-Myb involves recruitment of p300 and induction of histone acetylation followed by acetylation-induced chromatin dissociation. Epigenetics & Chromatin, 11, 35.

Publication 2018

anti-H3K27ac antibody (#

Anti antibody Anti c antibodies Cell line Chip Hybridoma Mouse Plasmids Primer Vector

Corresponding Organization :

Other organizations : University of Oslo

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Variable analysis

independent variables

Plasmids encoding TY-tagged c-Myb variants (empty vector, 3xTY1-c-Myb, 3xTY-c-Myb D152V)

dependent variables

Occupancy of the mim-1 locus (LECT2) determined with qRT-PCR

control variables

HD-11 cell line
ChIP analysis performed as previously described [53, 54]
Anti-Ty1 antibodies for IP of c-Myb
Anti-H3K27ac antibody for IP of H3K27ac

positive controls

Anti-Ty1 monoclonal mouse antibody produced from a hybridoma cell line [55]

negative controls

None explicitly mentioned

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