Injury induces HDAC5 phosphorylation and the formation of an HDAC5 gradient. Injury induces HDAC5 phosphorylation and the formation of an HDAC5 gradient. (A) Longitudinal sections of unligated or ligated sciatic nerve stained with p‐HDAC5 and the axon marker βIII tubulin (TUJ1). Colocalization of p‐HDAC5 with TUJ1 demonstrates HDAC5 axonal localization. (B) Sciatic nerves were treated with vehicle or the PKC inhibitor Gö6983 (1 mg/kg of weight) and ligated for 2 h. The level of p‐HDAC5 was analysed by western blot in ligated and unligated sciatic nerves. (C) Quantification of normalized p‐HDAC5 to total HDAC5 intensity (n=4, **P<0.01, mean±s.e.m.). (D) DRG neurons were axotomized at DIV7 (white dotted line) and stained with HDAC5, p‐HDAC5 and HDAC6 antibodies 3.5 h after axotomy. Bar, 100 μm. (E) Average intensity plot of HDAC5, p‐HDAC5 or HDAC6 with the corresponding average intensity plot of acetylated tubulin (n=6 for each; plot, mean±s.e.m.). (F) Cultured DIV7 DRG neurons were treated with or without ionomycin or Gö6983. HDAC5 was immunoprecipitated and analysed by western blot with anti‐kinesin‐1 and anti‐HDAC5 antibodies (TCL: total cell lysate prior to immunoprecipitation). (G) Sciatic and optic nerves were analysed by western blot for HDAC5. No HDAC5 can be detected in the optic nerve. Yongcheol Cho, and Valeria Cavalli EMBO J. 2012;31:3063-3078 © as stated in the article, figure or figure legend