Journal: Turkish Neurosurgery

Article Title: Protective effect of ad-vegf-bone mesenchymal stem cells on cerebral infarction

doi: 10.5137/1019-5149.jtn.11488-14.3

Figure Lengend Snippet: Figure 6: A) The immunohistochemical analysis showed that the reactive gliosis were attenuated after the treatment of BMSC- Ad-VEGF group (x200). The histogram showed the number of GFAP-positive cells. Compared with the other group, the number of GFAP- positive cells were lower after the treatment of BMSC- Ad-VEGF group (p<0.0001). Scale bar equals 100 μm. B) The expression of pAKT, AKT, pGSK3β and GFAP were detected by western blotting. The results showed that the proportion of pAkt/ Akt was increased on BMSC- Ad-VEGF group. There was no significant difference in the expression of pGSK3β and GFAP in 1 day. Otherwise, we founded that the expression of pGSK3β was increased, but the expression of GFAP were lower after the treatment of BMSC-Ad-VEGF group at 7, 14 days (P: PBS; B: BMSC- Ad; BV: BMSC-Ad-VEGF).

Article Snippet: Rat mesenchymal stem cell osteogenic differentiation medium (CYAGEN, CHINA), β-glycerophosphate, vitamin C, dexamethasone (Sigma), inverted phase contrast microscope (NIKON, Japan), medical clean bench (Jiangsu Sujing, China), adenovirus vector carrying human VEGF (Li Huiyong cooperative construction), stereotaxic instrument (Clinical Medical College of Jiangsu University), 2,3,5-3-triphenyl tetrazolium chloride (TTC, Amresco), 10% chloral hydrate, fishing line, anti-rat VEGF antibody (SANTA), anti-rat GFAP antibody (SANTA), anti-rat AKT antibody (Wuhan Boster), anti-rat pAKT antibody (Santa), rat GSK (Cell Signaling), and rat pGSK (Cell attracted increasing attention Signaling) were used.

Techniques: Immunohistochemical staining, Expressing, Western Blot

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: Three to four month-old HIVgp120tg and WT littermate male mice received intranasal recombinant murine IFNβ (50,000 U/25 g bodyweight) or vehicle once a week for four weeks and afterwards, the brains were assessed for neuronal damage and glial activation. Representative images of frontal cerebral cortex ( a ) and hippocampus ( b ) immunolabeled for neuronal synaptophysin (SYP; cortex layer III, hippocampus CA1) deconvolution microscopy; scale bar, 40 μm) and MAP-2 (cortex layer III (area between dashed lines) and hippocampus) fluorescence microscopy; scale bar, 100 μm), astrocytic GFAP (scale bar, 100 μm) and microglial Iba1 (red, DNA in blue, scale bar, 100 μm). ( c–f ) Quantification of microscopy data obtained in frontal cortex and hippocampus of sagittal brain sections of four to five month-old HIVgp120tg mice and controls treated with IFNβ or vehicle: ( c ) SYP in percent positive neuropil, ( d ) MAP-2 immunoreactivity as sum of fluorescence intensity (SFLI; arbitrary units); ( e ) fluorescence signal for astrocytic GFAP; ( f ) quantification of Iba1 + microglia (counts/microscopic field). Values are mean ± s.e.m.; ***p < 0.001, **p < 0.01, *p < 0.05; ANOVA and Fisher’s PLSD post hoc test; n = 4–5 animals per group/genotype.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Recombinant, Activation Assay, Immunolabeling, Microscopy, Fluorescence

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: Sagittal brains sections of HIVgp120tg and WT littermate mice previously treated with intranasal IFNβ or vehicle (veh) were immunolabeled for CCL4, neuronal MAP-2 or astrocytic GFAP. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was labeled with Hoechst (H) 33342. The fluorescence-labeled brain sections were analyzed using confocal laser-scanning microscopy. Representative images of cortex layer III are shown; scale bar, 50 μm.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Immunolabeling, Labeling, Fluorescence, Confocal Laser Scanning Microscopy

Journal: Journal of Neuroinflammation

Article Title: Sublethal enteroviral infection exacerbates disease progression in an ALS mouse model

doi: 10.1186/s12974-022-02380-7

Figure Lengend Snippet: Sublethal CVB3 infection triggers microglia/astrocyte activation and immune infiltration in mice. Representative images of GFAP, Iba1, CD19 and NK1.1 immunohistochemical staining in the hippocampus regions of the brain from mock- or CVB3-infected C57BL/6J or SOD1 G85R mice at day 10 PI ( A ) or week 60 PI (or humane endpoint) ( B ). Similar observation was made in the regions of infected cerebral cortex (data not shown). Images for other targets (CD68, CD79A, CD4, can CD8) can be found in the Additional file : Fig S2. Immune infiltrations were quantified by optical density based on the IHC staining in the hippocampus and cerebral cortex regions. Black boxes on the top right illustrate the enlarged images. Quantifications are presented as mean ± SEM ( n = 3–4 individual mouse brain samples for each target, timepoint and experimental condition). Scale bar = 100 μm. Statistical analysis was carried out using two-way ANOVA, followed by Bonferroni’s multiple comparison test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001

Article Snippet: Mouse brain tissues were immunostained using the follow primary antibodies diluted in Tris-buffered saline (pH 7.6) containing 1% bovine serum albumen: VP1 (1:2000 dilution, IgG2a monoclonal antibody, Mediagnost, Reutlingen, Germany), GFAP (1:200 dilution, mouse monoclonal antibody, StressMarq, Victoria, BC, Canada), Iba1 (1:200 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), TDP-43 (1:1000 dilution, rabbit polyclonal antibody, Proteintech, Rosemont, IL, USA), SQSTM1/p62 (1:200 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), ubiquitin (1:2000 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), CD68 (1:200 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), CD19 (1:500 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), CD79A (1:300 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), CD4 (1:500 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), CD8 (1:500 dilution, mouse monoclonal antibody, Santa Cruz Biotechnology, Dallas, TX, USA), and NK1.1 (1:1000 dilution, mouse monoclonal antibody, eBioscience, San Diego, CA, USA).

Techniques: Infection, Activation Assay, Immunohistochemical staining, Staining, Immunohistochemistry