H3 antibody ab1791

Cells were grown to mid-log phase at 32 °C in YES. For phosphate starvation experiments, cells in mid-log phase were washed twice in dH₂O before being grown in PMG (−PO₄) for 4 h. ChIP was performed essentially as described12 (link). Briefly, cells were fixed with 1% paraformaldehyde for 15 min at room temperature. Cells were lysed by bead beating (Biospec Prodcutes) and sonicated using a Bioruptor (Diagenode) sonicator at 5 °C on high for a total of 20 min (30 s ON/OFF cycles). Five microlitres of Rpb1 antibody (#2629; Cell Signaling), 2 μl GFP antibody (G10362; Life Technologies), 2 μl H3 antibody (ab1791; Abcam) and 1 μl of H3K9me2 antibody (m5.1.1; ref. 56 (link)) were used for IPs. RIP experiments were performed essentially as described13 (link). Hisx6-TEV-Protein A-tagged Mmi1 was captured from cell lysate with IgG Dynabeads (Life Technologies). Mmi1-bound RNA was isolated by phenol-chloroform extraction, DNase treated and reverse transcribed. Quantitative analysis was performed by qPCR.

GluR1 antibody (SC-55509), GluR2 antibody (SC-517265), STAT3 antibody (SC-482), pY705-STAT3 antibody (SC-7993), c-Myc antibody (SC-40), and HA-probe antibody (SC-7392) were from Santa Cruz Biotechnology (Santa Cruz). Acetylated-lysine antibody (9441), acK685-STAT3 antibody (2523), pS727-STAT3 antibody (9136), P70/85 S6 kinase antibody (9202), pERK1/2 antibody (9106), and ERK1/2 antibody (9102) were from Cell Signaling Technology (Boston). H3 antibody (ab1791) and VDAC1/porin antibody (ab14734) were from Abcam. Flag antibody (F9291) and tubulin antibody (T2200) were from Sigma.