Journal: Materials Today Bio

Article Title: Palladium–reduced graphene oxide nanocomposites enhance neurite outgrowth and protect neurons from Ishemic stroke

doi: 10.1016/j.mtbio.2024.101184

Figure Lengend Snippet: A) Schematic illustration of the synthetic process of Pd-RGO nanocomposite. B) Schematic diagram depicting the influence of palladium-reduced graphene oxide (Pd-RGO) nanocomposite on the outgrowth and repair of neurons. The length and complexity of neuronal neurites are enhanced significantly by Pd-RGO nanocomposites substrate that were possibly mediated by increasing of GAP-43, matrix proteins and electrons. C) Schematic diagram of Pd-RGO composites driving neuroprotection against ischemia/reperfusion injury in vivo in rodent model.

Article Snippet: After blocking the PVDF membrane with 5 % skim milk for 1 h at room temperature, primary antibodies were incubated at 4 °C for 15 h at 4 °C (rabbit polyclonal anti-MAP2, 1: 1000, Sigma; mouse monoclonal anti-β–III–tubulin, 1: 1000, Invitrogen; rabbit polyclonal anti-GAP-43, 1: 1000, Sigma; anti-GAPDH-HRP, 1 The membrane was incubated with HRP-conjugated secondary antibody at room temperature for 2 h after TBST washing.

Techniques: In Vivo

Journal: Materials Today Bio

Article Title: Palladium–reduced graphene oxide nanocomposites enhance neurite outgrowth and protect neurons from Ishemic stroke

doi: 10.1016/j.mtbio.2024.101184

Figure Lengend Snippet: Pd-RGO nanocomposites enhanced neuritic length and complexity of cultured neurons, compared with the RGO-treated group. A) Representative fluorescent images of DIV4 hippocampal neurons. Neurons were stained with Tau-1 (red) and MAP2 (green). Scale bar, 50 μm. B) Neurolucida tracings of representative Tau-1-immunolabeled neurons on day 4 after seeding. Scale bar, 50 μm. C–F) Quantification of total length of axons, axonal complexity index, number of axonal terminals and number of axonal nodes of neurons stained with axon-specific marker Tau-1 at DIV4. G) Western blot analysis of Tau-1 of hippocampal neurons on different groups and the relative optical densities ( n = 3). H) The representative fluorescence images of neurons on various substrates on day 7 after seeding, neurons were immune-stained with and MAP-2 (green), Scale bar represented 50 μm. I) Neurolucida tracings of representative MAP2-immunolabeled DIV7 neurons. Scale bar represented 50 μm. J-M) Quantification of total length of dendrites, average length of dendrites, number of dendritic nodes and dendritic complexity index of neurons on different groups. N) Quantification of dendritic complexity as measured by Sholl analysis. O) Western blot analysis of MAP2 and P) GAP-43 of hippocampal neurons on different groups. The experiments were repeated more than five times. All Data are expressed as mean ± SEM, n = 80–140 neurons in each group. Data were analyzed by one-way ANOVA, with post hoc Bonferroni post-tests; * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: After blocking the PVDF membrane with 5 % skim milk for 1 h at room temperature, primary antibodies were incubated at 4 °C for 15 h at 4 °C (rabbit polyclonal anti-MAP2, 1: 1000, Sigma; mouse monoclonal anti-β–III–tubulin, 1: 1000, Invitrogen; rabbit polyclonal anti-GAP-43, 1: 1000, Sigma; anti-GAPDH-HRP, 1 The membrane was incubated with HRP-conjugated secondary antibody at room temperature for 2 h after TBST washing.

Techniques: Cell Culture, Staining, Immunolabeling, Marker, Western Blot, Fluorescence

Journal: Materials Today Bio

Article Title: Palladium–reduced graphene oxide nanocomposites enhance neurite outgrowth and protect neurons from Ishemic stroke

doi: 10.1016/j.mtbio.2024.101184

Figure Lengend Snippet: The Pd-RGO nanocomposite significantly upregulated GAP-43 expression in cultured neurons. A) Representative images of primary hippocampal neurons at DIV7 immune-labeled with GAP-43 (red) and β-III tubulin (green), with counterstaining of nuclear marker Hoechst (blue). Scale bar, 30 μm. B, C) Western blot analysis (B) and the statistical results (C) of neuronal GAP-43 expression after Pd-RGO nanocomposite treatments at distinct concentrations at DIV7. Data are presented as means ± SEM. n = 3 for each group. * p < 0.05; One-way ANOVA with post hoc Dunnett's test.

Article Snippet: After blocking the PVDF membrane with 5 % skim milk for 1 h at room temperature, primary antibodies were incubated at 4 °C for 15 h at 4 °C (rabbit polyclonal anti-MAP2, 1: 1000, Sigma; mouse monoclonal anti-β–III–tubulin, 1: 1000, Invitrogen; rabbit polyclonal anti-GAP-43, 1: 1000, Sigma; anti-GAPDH-HRP, 1 The membrane was incubated with HRP-conjugated secondary antibody at room temperature for 2 h after TBST washing.

Techniques: Expressing, Cell Culture, Labeling, Marker, Western Blot

Journal: Materials Today Bio

Article Title: Palladium–reduced graphene oxide nanocomposites enhance neurite outgrowth and protect neurons from Ishemic stroke

doi: 10.1016/j.mtbio.2024.101184

Figure Lengend Snippet: Pd-RGO nanocomposite significantly facilitates dendritic outgrowth in cultured primary hippocampal neurons exposed to OGD/R. A) Representative fluorescence images of neurons exposed to OGD/R on various substrates on day 7 after seeding, neurons were immune-stained with Tau-1 (red) and MAP2 (green), Scale bar, 50 μm. And representative fluorescence images of neurons exposed to OGD/R on day 7 after seeding, neurons were immune-stained with GAP-43 (red) and β-III tubulin (green), Scale bar, 50 μm. B-E) Western blot analysis of MAP2, Tau-1 and GAP-43 in cultured hippocampal neurons exposed to OGD/R after Pd-RGO nanocomposite treated. n = 3–6 for each group. F) The representative fluorescence images of neurons exposed to OGD/R on the 10 μg mL −1 PDL-Pd-RGO nanocomposite-coated glass on day 7 after seeding, neurons were immune-stained with and MAP2 (green), Scale bar, 20 μm. G) Tracings of representative MAP2-immunolabeled neurons exposed to OGD/R on day 7 after seeding. Scale bar represented 50 μm. H–K) Quantification of total length of dendrites, average length of dendrites, number of dendritic nodes and dendritic complexity index of neurons exposed to OGD/R on the 10 μg mL −1 Pd-RGO nanocomposite-coated glass, which stained with dendritic marker MAP2 at DIV7. Data are expressed as mean ± SEM, n = 80–140 for each group, the experiment was repeated six times. L) Quantification of dendritic complexity as measured by Sholl analysis. All data are expressed as mean ± SEM, n = 80–140 for each group. Data were analyzed by one-way ANOVA, with post hoc Bonferroni tests. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: After blocking the PVDF membrane with 5 % skim milk for 1 h at room temperature, primary antibodies were incubated at 4 °C for 15 h at 4 °C (rabbit polyclonal anti-MAP2, 1: 1000, Sigma; mouse monoclonal anti-β–III–tubulin, 1: 1000, Invitrogen; rabbit polyclonal anti-GAP-43, 1: 1000, Sigma; anti-GAPDH-HRP, 1 The membrane was incubated with HRP-conjugated secondary antibody at room temperature for 2 h after TBST washing.

Techniques: Cell Culture, Fluorescence, Staining, Western Blot, Immunolabeling, Marker

Journal: PLoS ONE

Article Title: A Peptide Mimetic Targeting Trans -Homophilic NCAM Binding Sites Promotes Spatial Learning and Neural Plasticity in the Hippocampus

doi: 10.1371/journal.pone.0023433

Figure Lengend Snippet: (a) 125,000 cells/cm 2 CGNs were left to differentiate for six days in the presence of high potassium (40 mM), followed by a 48 h incubation period with or without 15 nM EndoN. Cultures were double immunostained for GAP-43 (green) and PSA-NCAM (red). 12,500 cells/cm 2 Hippocampal neurons (b) and CGNs (c) were grown for 24 h in the presence or absence of 1.74 µM plannexin and treated with 30 nM EndoN ( b ) or 15 nM EndoN ( c ). Results from 4–5 experiments are expressed as a percentage ± SEM, with unstimulated controls set at 100%. *** p <0.001 vs. untreated controls; +++ p <0.001 vs. peptide-treated cultures without EndoN treatment.

Article Snippet: Polyclonal rabbit anti-GAP-43 antibodies were obtained from Chemicon (Temecula, CA, USA).

Techniques: Incubation