Anti hdac9

Immunohistochemical analyses were performed on LNAT, LT and LP tissue samples fixed with formaldehyde 10% or Hope I/II reactive Method (Innovative Diagnostik-System, Germany), and subsequently using 1–2 micrograms of specific antibodies anti-MEOX2, anti-TWIST1, anti-HDAC9, and anti-EVX1 (Santa Cruz Biotechnology, Dallas, Texas, USA), by incubation in a humidity chamber and in addition of the antigen-specific reaction with avidin-biotin peroxidase/diamino-benzidine system (Dako, Carpinteria, CA, USA). A Ventana System device (Roche, Tucson, Arizona, USA) and a transmitted light microscopy study (Leica, Bannockburn, IL, USA), were used to obtain photomicrographs at 40X and 80X magnification.

The following antibodies were used: anti‐α‐tubulin (sc‐8035), anti‐Sp1 (sc‐420), anti‐HDAC1 (sc‐81598), anti‐HDAC2 (sc‐9959), anti‐HDAC4 (sc‐46672), anti‐HDAC5 (sc‐133225), anti‐HDAC7 (sc‐74563), anti‐HDAC8 (sc‐17778), anti‐HDAC9 (sc‐398003), anti‐HDAC10 (sc‐393417), and anti‐PKCζ (sc‐17781; Santa Cruz Biotechnology) antibody; anti‐phospho ser/thr (#96315), anti‐active caspase‐3 (#9664), and acetyl‐histone 4 (#8647S; Cell Signaling) antibody. Rabbit polyclonal anti‐PLD antibody that recognizes both PLD1 and PLD2 was generated as described previously (Min et al., 2001). The signal densities on the blots were measured with ImageJ (Wayne Rasband) and normalized using anti‐α‐tubulin antibody.