Tat cn21

Drugs (Sigma, µM) were AP5 (100), AMPA (10), bicuculline (10), CsCl (2 mM), KA (20), ketanserin (1), KN-93 (1), MK-801 (50), NiCl₂ (50), nifedipine (50–75), NMDA (30 or 50), tatCN21 and tatCtrl (15, synthesized by K.U.B.), and TBOA (100, Tocris). Drugs were dissolved in ACSF (0.001% DMSO as needed) and tatCN21 and tatCtrl were dissolved in water before into the internal electrode solution.

Chemical iLTP was induced in DIV14–18 hippocampal neurons via bath application of 20 μM NMDA and 10 μM CNQX in HBS solution (145 mM NaCl, 2 mM KCl, 10 mM HEPES, 2 mM CaCl₂, 2 mM MgCl₂ 10 mM glucose, pH 7.4) for 2 min at 37°C as previously described (Petrini et al., 2014 (link)). Neurons were harvested or fixed at specific time points post-stimulation as stated in the figures and/or figure legends. Cycloheximide was supplemented into the bath media and conditioned media at 10 μg/ml and left in the culture media until the neurons were ready to be harvested or fixed. Cyclosporin A (Tocris), FK506 (Tocris) and TAT-CN21 (kind gift from Dr. Ulli Bayer) were used at 5 μM and added to cell culture media 15 minutes prior to iLTP induction and left in the culture media following stimulation until the neurons were ready to be harvested. Likewise, BAPTA-AM (Tocris) was used at 20 μM and Nimodipine at 10 μM. Neurons were treated with Trichostatin A (TSA) for 16 hours at 1 μM prior to iLTP induction.