H3k18ac

His-tagged fusion proteins were prepared using Ni-NTA agarose beads (Qiagen, UK) according to the manufacturer’s protocol and HA epitope-tagged Pkc1p were prepared essentially as described previously (Darieva et al., 2012 (link)). Recombinant histone H3 was made commercially (Active Motif).
Pulldown assays with immunoprecipitated HA-Pkc1 from DZ2 cells and in vitro translated Rtt109 were carried out as described previously (Pic-Taylor et al., 2004 (link)).
To detect epitope-tagged derivatives by western analysis, anti-HA (Roche) and anti-tubulin TAT-1 (CRUK), H3K56ac (Active motif), H3K9ac (Abcam), H3K18ac (Abcam), H3 (Active motif), myc epitope (Santa Cruz), H3T45-P (Active motif) antibodies and Supersignal west dura substrate (Pierce) were used.
Rabbit polyclonal monospecific antibody against the phosphothreonine at amino acid 46 on yeast Rtt109 (Rtt109 T46-P) was generated from immunizing rabbits with a KLH-conjugated peptide (H-DDKRVPKST(PO₃H₂)IKTC-NH2), and then cross-affinity purification of polyclonal antibodies specific to modified Rtt109 T46-P or non-modified pRtt109 was performed (Eurogentec). For immunoblotting analysis we used 1:1000 dilution for both antibodies.

Cell culture reagents were purchased from Gibco. Antibodies and reagents used were as follows: Cav-2 (BD 610685), lamin A/C (BD 612162) and E-cadherin (BD 610182), antibodies from BD Transduction Laboratories; lamin A/C (sc-20681), lamin B1 (sc-30264), α-tubulin (sc-5286), maltose-binding protein (MBP) (sc-73416), emerin (sc-15378), GFP (sc-9996), histone H3 (sc-10890), histone H1 (sc-8030), protein-tyrosine phosphatase 1B (PTP1B) (sc-1718), GCN5 (sc-20698) and p300 (sc-585) antibodies from Santa Cruz Biotechnology; pY19-Cav-2 (ab3417), lamin B-receptor (LBR) (ab169306), H3K9me3 (ab8898), H3K9ac (ab32129), H3K18ac (ab1191) and H3K27ac (ab4729) antibodies from Abcam; GFP (#2555) antibody from Cell Signaling; AcH3 (06-599), RNA Pol II (05-623), H3K4ac (07-539), and H3K14ac (07-353) antibodies from Millipore; FITC-conjugated anti-mouse (F9006), TRITC-conjugated anti-rabbit (T5268), horseradish peroxidase (HRP)-conjugated anti-mouse (A4416) and anti-rabbit (A6154) antibodies and 4′-6-diamidino-2-phenylindole (DAPI) (D8417), curcumin (C1386), butyrolactone 3 (M2449) and sodium ortho-vanadate (S6508) from Sigma; human insulin from Eli Lilly.

Histone modifications primary antibodies: H3K4me3 (Millipore, Billerica, MA), H3K9ac (Cell Signaling, Boston, MA), ChIP grade H3K9ac (Abcam, UK), H3K9ac (Millipore, Billerica, MA), H3K9me3 (Millipore), H3K14ac (Millipore), H3K18ac (Cell Signaling), H3K18ac (Abcam), ChIP grade H3K27ac (Abcam), H3K27me3 (Millipore), H3 (Abcam), H4K16ac (Santa Cruz Biotechnology Inc., Dallas, Texas).
Non-histone primary antibodies: HA (Abcam).

The CellTiter 96 Aqueous ONE Solution Cell Proliferation Assay was purchased from Promega Corp. (Madison, WI, USA). NVP-AUY922 (AUY922) (>99% purity) was from Selleckchem.com (Houston, TX, USA). Ganetespib (STA9090) (98.79% purity) and SNX2112 (98% purity) were purchased from ApexBio (Houston, TX, USA). AT13387 (>98%) was from MedChem Express (Princeton, NJ, USA), CUDC305 (>98% purity) was from AbMole BioScience (Houston, TX, USA), and dimethyl sulfoxide (DMSO) was from Sigma-Aldrich (St. Louis, MO, USA). AUY922, ganetespib, SNX2112, AT13387, and CUDC305 were dissolved in DMSO separately and stored at −20 °C. Polyclonal antibodies against histone H3, H4, H3K4me3, H3K18ac, and H4K12ac were purchased from Abcam (Cambridge, MA, USA). Monoclonal or polyclonal antibodies against acetyl-histone H3, acetyl-histone H4, and H3K27ac were bought from EMD Millipore Corporation (Billerica, MA, USA). Monoclonal or polyclonal antibodies against histone H3K4me1, H3K4me2, H3K23ac, H3K79me1, H3K79me2, H4K8ac, and H4K20me3 were obtained from Cell Signaling Technology (Danvers, MA, USA). Restore Western Blot Stripping Buffer was from Thermo Scientific (Rockford, IL, USA). All other reagents were from Sigma-Aldrich.