Poly l lysine hydrobromide p1274

Native BoNT/C and BoNT/A1 were purified as previously described [66 (link), 67 (link)]. Cytosine β-D-arabinofuranoside hydrochloride (C6645), DNAse I from bovine pancreas (DN25), poly-L-lysine hydrobromide (P1274) and trypsin (T4799) were from Sigma Aldrich. μ-Conotoxin GIIIB is from Alomone, Jerusalem, Israel. Primary antibodies: anti-SNAP-25 (SMI81, ab24737) was from Abcam. Anti-SNAP-25 (cleaved) and syntaxin-1A/1B polyclonal antibodies were produced in our laboratory and previously characterized [37 (link), 68 (link)]. Secondary antibodies conjugated to HRP were from Calbiochem; secondary antibodies for immunofluorescence conjugated to Alexa Fluorophores 488 or 555 and α-Bungarotoxin conjugated to Alexa 647 were from Thermo Scientific, Waltham, MA, USA.

High-glucose and glucose-free Dulbecco’s modified eagle medium (DMEM), neurobasal medium, B27, MitoSOX™ Red mitochondrial superoxide indicator, Alexa Fluor 488 and 532 labeled fluorescent secondary antibodies, BCA Protein Assay Kit and other reagents were purchased from Thermo Fisher Scientific Inc (ThermoFisher Scientific, Waltham, MA, USA). Poly-L-lysine hydrobromide (P1274) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The rabbit anti-AIF antibody (#4642) and rabbit anti-PARP antibody (#9542) were purchased from Cell Signaling Technology (Danvers, MA, USA). The mouse anti-MAP2 antibody (ab221693) was purchased from Abcam (Abcam, Cambridge, UK). The mouse anti-β-actin antibody (sc-47778) and mouse anti-pADPr antibody (sc-56198) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The RIPA lysate was purchased from Kangwei Century Biotechnology (Cwbio, Beijing, China). The Tunel assay kit (G1501) was purchased from Wuhan Servicebio Company (Wuhan, China). The DCFH-DA kit was purchased from Beyotime (Beijing, China). The MCAO thread plug was purchased from RWD life science Co., Ltd. (Shenzhen, China). Other chemical reagents were analytically pure.