Anti atf3

Lumbar DRG-sciatic nerve/central branch explants were dissected as described above (see Section 2.5). Following 3 h of 10% mechanical stretch, DRG were dissected from their respective nerves and fixed with 4% PFA for 4 h at RT. Then, DRG were washed once with 1× PBS and then kept in 30% sucrose solution for 24 h at +4 °C. DRG were cryosected in 18 μm-thick sections. Sections were permeabilized in 0.25% TritonX-100/1× TBS solution and blocked in 5% donkey serum/1× TBS solution for 2 h at RT. Sections were incubated o/n with the primary antibody, anti-ATF3 (Activating Transcription Factor 3, 1:200, rabbit, Novus Biologicals, NBP1-85816, Littleton, CO, USA), anti-H3K9K14ac (Histone 3 lysine 9 lysine 14 acetylation, 1:1000, rabbit, Sigma Aldrich, ZRB06599, St. Louis, MO, USA), and β-III-tubulin (1:2000, mouse, Promega GmbH, G7121, Walldorf, Germany), diluted in 0.25% TritonX-100/1% donkey serum in 1× TBS solution. DRG were washed three times with 1× TBS and then incubated for 1 h at RT with the secondary antibody, AlexaFluor-594 (1:1000, donkey anti-rabbit, Life Technologies, A21207, Carlsbad, CA, USA), AlexaFluor-488 (1:1000, donkey anti-mouse, Life Technologies, A21202, ThermoFischer, MA, USA). DRG were washed three times with 1× TBS and counterstained with DAPI for 15 min. Sections were washed once with 1× TBS and coverslipped with Fluoromount-G.