Journal: Scientific Reports

Article Title: Stilbene derivatives promote Ago2-dependent tumour-suppressive microRNA activity

doi: 10.1038/srep00314

Figure Lengend Snippet: (a) MDA-MB231-luc-D3H2LN cells (200 cells) were injected into the mammary fat pad of six-week-old female SCID hairless outbred mice (n = 5). They were then treated with resveratrol (25 mg/kg/day) by intraperitoneal injection every day for 8 days. Tumour growth was monitored by injecting luciferin in the mice followed by measuring bioluminescence using an IVIS imaging system. Representative mouse images at day 8 (upper panel) and quantified bioluminescence images at day 8 (lower panel) are shown. (b) MDA-MB231-luc-D3H2LN cells were treated with resveratrol or DMSO (control) at the specified doses for 3 days. The percentage of CD44 + /CD24 − cells after compound treatment in independent experiments with MDA-MB231-luc-D3H2LN cell populations is shown. The CD44 + /CD24 − denoting the CSC-enriched fraction. (c) MDA-MB231-luc-D3H2LN cells were grown, treated with resveratrol or DMSO (control) for 1 day, and then subjected to an invasion assay. Representative photographs (upper panel) and quantification (lower panel) are shown. Scale bar: 100 μm. (d) The miR-141 and miR-200c expression levels in MDA-MB231-luc-D3H2LN cells. The expression levels of the indicated miRNAs were examined in MDA-MB231-luc-D3H2LN cells after 48 hour resveratrol treatment (all data are shown as the mean ± s.e.m., *P<0.05, **P<0.01, ***P<0.001).

Article Snippet: At least one million cells were pelleted by centrifugation at 180 x g for 5 minutes at 4°C, resuspended in a 5-μL mixture of a monoclonal mouse anti-human CD44-FITC antibody (Becton Dickinson, clone L178) and a monoclonal mouse anti-human CD24-APC antibody (Biolegend, clone ML5), and incubated for 30 minutes at 4°C.

Techniques: Injection, Imaging, Control, Invasion Assay, Expressing

Journal: Journal of Neuroinflammation

Article Title: Genetically enhancing the expression of chemokine domain of CX 3 CL1 fails to prevent tau pathology in mouse models of tauopathy

doi: 10.1186/s12974-018-1310-6

Figure Lengend Snippet: LPS-induced tau phosphorylation and microglial activation are exacerbated in Cx 3 cl1 105Δ mice. a – d Two-month-old fractalkine ( Cx 3 cl1 −/− )-deficient mice and the mice exclusively expressing the chemokine domain (lacking the mucin-like domain, red) (CX 3 CL1 105Δ ) with a Myc tag were injected with LPS (3 mg/kg b.w; i.p) or vehicle (VEH, Hank’s balanced salt solution or HBSS) and sacrificed 24 h post-injection. e – f Western blotting of the hippocampi revealed significantly increased total tau (Tau5) (> 1.5-fold) in VEH-treated Cx 3 cl1 105Δ vs. Cx 3 cl1 −/− mice (mean + SEM; ** p < 0.01; n = 3; two-way ANOVA followed by Tukey’s post hoc test). Both AT8/Tau5 and AT180/Tau5 ratios were significantly higher in LPS-treated Cx 3 cl1 105Δ compared to LPS-treated Cx 3 cl1 −/− or Non-Tg mice (mean + SEM; * p < 0.05; ** p < 0.01; n = 3; two-way ANOVA with Tukey’s post hoc test). g Immunohistochemistry (IHC) analysis revealing a modest increase in AT8 (pS199/pS202 tau) among experimental genotypes or between VEH- or LPS-injected mice in the CA3 hippocampal areas. Scale bar, 20 μm. h – k IHC showing elevated Iba1 + /F4/80 + reactive microglia in VEH-treated Cx 3 cl1 105Δ mice that is enhanced with LPS treatment. Quantification reveals statistically higher form factor units (higher number means more towards circular contour) for Iba1 + microglia in Non-Tg and Cx 3 cl1 105Δ mice in LPS-treated groups (mean + SEM; *** p < 0.0001 vs. ** p < 0.01 for Non-Tg with LPS; two-way ANOVA with Tukey’s post hoc test; n = 3–6 mice per group). Scale bars ( h , j ) 25 μm

Article Snippet: The sections were incubated overnight with AT8, AT180, or myeloid cell surface markers (CD45 #MCA1388 1:500; BioRad; F4/80 #MCA497G 1:500 BioRad; Iba1 #019-19741 1:500; Waco) and respective biotinylated secondary antibodies (Vector Laboratories Catalog # BA-2000, BA-9400, BA-1000) and developed using 3-3′-diaminobenzidine with or without the nickel enhancer.

Techniques: Phospho-proteomics, Activation Assay, Expressing, Injection, Western Blot, Immunohistochemistry