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