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Fig. 1 | Molecular Brain

Fig. 1

From: Helix-A peptide prevents gp120-mediated neuronal loss

Fig. 1

The neuroprotective effect of peptides against gp120 depends on their binding to MTs. a. Helix-A and other peptides were purchased from Genscript, Piscataway, NJ. The binding affinity of peptides to assembled MTs (Cytoskeleton Inc., Denver, CO), expressed as KD, was determined by surface plasmon resonance (Biacore™ T200, GE Health care Bio-Science, Piscataway, NJ) as previously described [8]. b-c. Rat cortical cultures were prepared from embryonic day 14–15 Sprague Dawley rats (Taconic, Derwood, MD) as previously described [8]. Neurons were exposed to gp120ADA (5 nM) (Immunodiagnostics Inc., Woburn, MA) alone or in combination with Helix-A MSN or the other four modified peptides crosslinked to MSNs (5 μM each), for 24 h. Neurite length (a) and cell survival (b) were then determined as previously described [8]. Boiled gp120ADA was used as a control. b. Example of images of neurons (20×) stained with a mouse MAP2 antibody (1:5000; MilliporeSigma, St Louis, MO) after exposure to boiled gp120 (control) or gp120ADA for 24 h. Neurite lengths were determined by analyzing MAP2 immunoreactivity in three randomly selected fields (10 neurons per field) from three separate experiments using ImageJ, as detailed elsewere [8]. *p < 0.0001 vs control, **p < 0.0001 vs gp120; One-way ANOVA followed by multiple comparisons by Tukey’s test. c. Neuronal survival was determined by Hoechst/PI staining (MilliporeSigma) as previously described [8]. Results are expressed as mean ± SEM (from 3 separate experiments with 2 duplicates per condition). #p < 0.005 vs control, ##p < 0.0004 vs gp120; One-way ANOVA followed by multiple comparisons by Tukey’s test

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