Journal: Neural Regeneration Research

Article Title: Small molecule inhibitor DDQ-treated hippocampal neuronal cells show improved neurite outgrowth and synaptic branching

doi: 10.4103/NRR.NRR-D-24-00157

Figure Lengend Snippet: Antibodies used in western blot analysis

Article Snippet: Synaptophysin , Rabbit polyclonal 1:500 , NBP2-25170 , Novus Biological, Littleton, CO, USA , , Anti-rabbit polyclonal IgG 1:1000 , 35552 , Thermo Fisher Scientific.

Techniques: Western Blot

Journal: Neural Regeneration Research

Article Title: Small molecule inhibitor DDQ-treated hippocampal neuronal cells show improved neurite outgrowth and synaptic branching

doi: 10.4103/NRR.NRR-D-24-00157

Figure Lengend Snippet: Antibodies used in immunofluorescence analysis

Article Snippet: Synaptophysin , Rabbit polyclonal 1:500 , NBP2-25170 , Novus Biological, Littleton, CO, USA , , Anti-rabbit polyclonal IgG 1:1000 , 35552 , Thermo Fisher Scientific.

Techniques: Immunofluorescence

Journal: Journal of Neurovirology

Article Title: Antiretroviral drugs induce oxidative stress and neuronal damage in the central nervous system

doi: 10.1007/s13365-013-0227-1

Figure Lengend Snippet: Antiretroviral drugs induce neuronal damage in vivo. a – d Formalin-fixed, paraffin-embedded tissue sections from hippocampus of pig-tailed macaques that were either uninfected ( n = 6), SIV infected but not cART treated ( n = 7), or SIV infected and treated with cART (tenofovir, atazanavir, saquinavir, and L-870812a; n = 4) were prepared for immunofluorescent analysis and were triple labeled for MAP2 ( red ), synaptophysin ( green ), and GFAP. Sections were visualized by laser confocal microscopy and images were quantified for MAP2, synaptophysin and GFAP expression. a Representative composite images of two cases per group which were stained with MAP2 and synaptophysin are shown. Scale bar = 30 μm. b Quantification shows the resolution of GFAP immunoreactivity in SIV(+)/cART group, compared with SIV(+)/placebo group (one-way ANOVA, * p < 0.05). No changes were observed in MAP2 expression between groups ( c ), but there were statistically significant decreases in synaptophysin immunoreactivity ( d ) in SIV(+)/cART group, as compared with SIV(+)/untreated and uninfected groups (one-way ANOVA, * p < 0.05, ns not significant). e , f Fresh-frozen tissue sections from the frontal cortex of pig-tailed macaques that were either uninfected ( n = 3), SIV infected but not cART treated ( n = 6), or SIV infected and cART treated ( n = 6) were used for standard protein extraction and subsequent immunoblotting for the expression of CaMKII. Actin was used as a loading control. A representative immunoblot is shown. Quantification shows statistically significant decreases in CaMKII in the cART-treated group, as compared with the uninfected group or the SIV(+)/untreated group (one-way ANOVA, * p < 0.05). g Whole cell lysates prepared from hippocampus of rats treated for 7 days with AZT/Rit/Saq ( n = 4) or vehicle ( n = 2) were immunoblotted for synaptophysin and MAP2. A band from the coomassie blue staining is included to control for equal loading and protein degradation

Article Snippet: Chemicals and reagents comprise the following: (1) AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH, antiretroviral reagents; (2) Abcam (Cambridge, MA), mouse monoclonal NAD(P)H/quinone oxidoreductase-1 (NQO-1) antibody (A180) and mouse monoclonal synaptophysin antibody (SY38); (3) BioRad (Hercules, CA), Biosafe Coomassie stain, immunoblot polyvinylidene fluoride (PVDF) membrane and prestained broad range molecular weight ladder; (4) Cell Signaling Technology (Danvers, MA), rabbit polyclonal antibody raised against cleaved caspase-3; (5) Citifluor, Ltd. (London, UK), citifluor AF1.

Techniques: In Vivo, Formalin-fixed Paraffin-Embedded, Infection, Labeling, Confocal Microscopy, Expressing, Staining, Protein Extraction, Western Blot, Control

Journal: Journal of Neurovirology

Article Title: Antiretroviral drugs induce oxidative stress and neuronal damage in the central nervous system

doi: 10.1007/s13365-013-0227-1

Figure Lengend Snippet: Therapeutically relevant combination antiretroviral drug treatments are neurotoxic in vitro. a – c Primary rat cortical neuroglial cultures aged 21 days in vitro ( DIV ) on coverslips were exposed to Rit ( a ), Saq ( b ), or AZT ( c ) at increasing doses for 48 h, followed by hand counting for MAP2-positive cells ( n = 3; vehicle, 0.04 % DMSO; * p < 0.05, one-way ANOVA with post-hoc Newman–Keuls). d – f 21DIV primary neuroglial cultures grown in 96-well plates were treated with increasing doses of Rit ( d ), Saq ( e ), or AZT ( f ) for 48 h, followed by MAP2 cell-based ELISA ( n = 2; vehicle, 0.04 % DMSO; * p < 0.05, one-way ANOVA with post-hoc Newman–Keuls). g Primary neuroglial cultures were treated with AZT (25 μM), Rit (10 μM), or Saq (1 μM) at day zero. Ninety percent of the media was changed with conditioned media supplemented with a fresh drug stock every 2 days, and cultures were analyzed by MAP2 cell-based ELISA at days 4 and 8 ( n = 2; vehicle, 0.04 % DMSO; * p < 0.05, one-way ANOVA with post-hoc Newman–Keuls). h , i Cultures grown on coverslips were exposed to the indicated treatments and synaptophysin-positive puncta were determined ( n = 3; vehicle, 0.04 % DMSO; * p < 0.05, one-way ANOVA with post-hoc Newman–Keuls). j , k Primary neuroglial cultures were exposed to the indicated drug combinations (AZT, 25 μm; Rit, 10 μm; Saq, 1 μm) for 48 h, followed by hand counting for MAP2-positive cells ( j ) or MAP2 cell-based ELISA ( k ) ( n = 2; vehicle,: 0.04 % DMSO; * p < 0.05; # p < 0.01, one-way ANOVA with post-hoc Newman–Keuls). l Primary neuroglial cultures that were exposed to the indicated drug combinations (AZT, 25 μm; Rit, 10 μm; Saq, 1 μm) for 16 h were analyzed for synaptophysin-positive puncta ( n = 2; vehicle, 0.04 % DMSO; * p < 0.05, one-way ANOVA with post-hoc Newman–Keuls)

Article Snippet: Chemicals and reagents comprise the following: (1) AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH, antiretroviral reagents; (2) Abcam (Cambridge, MA), mouse monoclonal NAD(P)H/quinone oxidoreductase-1 (NQO-1) antibody (A180) and mouse monoclonal synaptophysin antibody (SY38); (3) BioRad (Hercules, CA), Biosafe Coomassie stain, immunoblot polyvinylidene fluoride (PVDF) membrane and prestained broad range molecular weight ladder; (4) Cell Signaling Technology (Danvers, MA), rabbit polyclonal antibody raised against cleaved caspase-3; (5) Citifluor, Ltd. (London, UK), citifluor AF1.

Techniques: In Vitro, In-Cell ELISA