Zymo spin chip kit

Cells growing in 10 or 15-cm plates were cultured in RPMI 1640 (ATCC, 30-2001) supplemented with 5% (v/v) charcoal stripped fetal bovine serum (CSS) (Gibco, A33821) for 48 h. Plates were then treated in biological replicates with vehicle, 5 μM MDV or 5 μM MDV + 10 μM Enzastaurin for 24 h. Samples were subsequently processed using the Zymo-Spin ChIP Kit (D5209) and either a H3T6ph antibody (Abcam, ab222768), H3K4Me2 (Cell Signaling Technology, 9725), H3K4Me1 (Cell Signaling Technology, 5326), LSD1 (Abcam, 129195) or a rabbit IgG antibody (Cell Signaling Technology, 2729). The precipitated DNA was evaluated by qRT-PCR using the Maxima SYBR Green qPCR Master Mix (Life Technologies) on a Bio-Rad CFX Touch Real-Time PCR system according to manufacturer instructions. Data is reported as percent of input. Primer sequences are as follows: ARBS2d’ forward: GCT CAG AGA GGT TTT AGT TGT G, ARBS2d’ reverse: CAA AAT GTC TAA GCT GGA AGC AC; ARBS2b’ forward: GTC TTG CTT TCC TAG AAG GTG AC; ARBS2b’ reverse: CAA GGA GAA AAT CTG AGT CCT GAG; ARBS2b forward: CAC ATG GAG TGC TGT TTG GT, ARBS2b reverse: GTA AAC ATC AGT GAG GAT GGT G; ARBS2e forward: GCA GAG AGT TTT TGG TGC ATA TC, ARBS2e reverse: CAA AGA TAC CTG ATG AAG GCT CTG; ARBS2g forward: CAG ACT TTA GAT TTA GGG GTT GG, ARBS2g reverse: GTC TAT GGC TGC TTT CAT CCT AC.