Journal: Cell Reports Methods

Article Title: Patterning ganglionic eminences in developing human brain organoids using a morphogen-gradient-inducing device

doi: 10.1016/j.crmeth.2023.100689

Figure Lengend Snippet:

Article Snippet: XAV939 [5 μM] , BioGems , 2848932-25mg.

Techniques: Recombinant, Gene Expression, Software, Imaging

Journal: bioRxiv

Article Title: Loss of the Wnt/β-catenin pathway in microglia of the developing brain drives pro-inflammatory activation leading to white matter injury

doi: 10.1101/334359

Figure Lengend Snippet: Analysis of microarray data from MACS purified CD11B+ MG/Mφ from IL-1β and PBS administered mice was undertaken. This revealed ( a ) that changes in the Wnt signalling pathway are strongly significantly associated with MG/Mφ activation, see also Supp Figure 2 . In ( b ) a cohesive network of multi-modal interactions between Wnt pathway genes in the MG/Mφ co-expression network can be generated via analysis using the GeneMania tool network . Annotation of this network with a bold red circle indicates that the gene is down-regulated in the microarray data at P5 and P10, genes with a finer red line are down-regulated at a single time point and those in purple display a switch, being significantly higher at one and significantly lower at the other time point. In ( c ) validation of robust down-regulation in MG/Mφ of mRNA expression of Ctnnb1, Tcf1, Lef1 and Fzd 4,6 mRNA by RT-qPCR in CD11B+ MG/Mφ MACSed from brains of PBSor IL-1β-exposed mice at P1, 2, 3, 5 and 10. See also Supp Figure 3 . Analysis made with a two-way ANOVA with post hoc Bonferroni‘s test: * p<0.05, ** p<0.01 *** p<0.001, numbers of samples for each data point shown on the graphs in corresponding colours. In ( d ) quantification of β-catenin by ELISA in CD11B+ MG/Mφ MACSed from brains of PBS or IL-1β-exposed mice at P3. β-catenin protein levels were reduced and are presented as a fold change relative to PBS-injected mice, (mean±SEM). Analysis made with a Student‘s t-test, * p<0.05. In ( e ) XAV939, a β-catenin pathway inhibitor induced a pro-inflammatory MG activation as assessed with quantification of Nos2, Ptgs2, Il1rn and Socs3 mRNA by RT-qPCR in primary MG exposed to XAV939 (1µM) or DMSO for 4 hours. mRNA levels are presented as a fold change relative to DMSO group, mean±SEM, Student‘s t-test: * p<0.05, ** p<0.01. In ( f ) a reductions of pro-inflammatory MG activation by gene silencing using siRNA for Axin2 (30µM), a β-catenin inhibitor, or scrambled control (Ctrl) siRNA transfected into MG for 48 hours prior to 4 hours of IL-1β exposure, see also Supp Figure 3h. siRNA for Axin2 reducesIL-1β-induced Nos2, Ptgs2, Il1rn and Socs3 mRNA. Data are presented as a fold change relative to negative siRNA group, mean±SEM, one-way ANOVA with post hoc Newman-Keuls‘s test: effects of IL-1β on gene expression are shown with + p<0.05, ++ p<0.01, +++p<0.001; effects of Axin2 siRNAto alter the IL-1β effects are shown with * p<0.05, ** p<0.01, ***p<0.001. In ( g ) Inhibition of GSK3β, aβ-catenin inhibitor, by CT99021 (1µM) treatment 1 hour prior to 4 hours of IL-1β exposure reduces IL-1β-induced Nos2, Ptgs2, Il1rn and Socs3 mRNA overexpression. mRNA levels are quantified by qRT-PCR and presented as a fold change relative to control/DMSO group, (mean±SEM, One-way ANOVA with post hoc Newman-Keuls‘s test: + p<0.05, ++ p<0.01, +++p<0.001 indicate a statistically significant difference between PBS+DMSO and IL-1β+DMSO; * p<0.05, ** p<0.01, ***p<0.001. In ( h )effects of modulating Wnt with XAV939, Axin2 siRNA and CT99021 on phosphorylation of β-catenin(PSer45 β-catenin/β-catenin ratio) were quantified by ELISA in MG. XAV939 (1µM) inhibits β-catenin while CT99021 (1µM) or siRNA against Axin2 (30µM) activate it when you compare to their respective vehicle (DMSO or negative control siRNA). Data are expressed as the mean±SEM (*p<0.05 in a Mann-Whitney test). For all bar graphs the n for each data point is indicated on or above the bar.

Article Snippet: To inhibit the Wnt/β-catenin pathway, we used XAV939 (Sigma-Aldrich, France).

Techniques: Microarray, Purification, Activation Assay, Expressing, Generated, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Injection, Transfection, Inhibition, Over Expression, Negative Control, MANN-WHITNEY

Journal: bioRxiv

Article Title: Loss of the Wnt/β-catenin pathway in microglia of the developing brain drives pro-inflammatory activation leading to white matter injury

doi: 10.1101/334359

Figure Lengend Snippet: Wnt signalling was modulated in vivo with pharmacological techniques in zebrafish ( a-c ) and mice ( d ) and using gene knockout in mice ( e-g ). In ( a ) LPS microinjection (5ng/injection) into the zebrafish hindbrain at 72 hpf in pu1::Gal4-UAS::TagRFP morpholinos followed by analysis at 120 hpf. In ( b ) quantification of RFP labelled MG and in ( c ) representative images of RFP MG in these zebrafish brains after LPS injection in the presence or not in their growth solution of modulators of Wnt/β-catenin signalling, LiCl (80mM), XAV939 (5μM), CT99021 (3μM). Agonists of Wnt (LiCl, CT99021) prevented LPS driven MG immunoreactivity and further antagonizing Wnt (XAV939) enhanced this inflammatory reactivity. Data are mean±SEM, One-way ANOVA with post hoc Newman-Keuls‘s test:** p<0.01, ***p<0.001. Scale bar: 40μm. In ( d ) analysis of CD11B+ cells isolated from P1 mice injected i.c.v. with XAV939 (0.5 nmol) alone demonstrating increased MG/Mφ activation. Quantification of Nos2, Ptgs2, Tnf α, Il1rn, Socs3 and Il4ra mRNA by RT-qPCR. mRNA levels are presented as a fold change relative to PBS/DMSO i.c.v. injected mice. Data expressed as the mean±SEM, Student‘s t-test: * p<0.05, ** p<0.01 and ***p<0.001. In (e-g) β-catenin deficit in MG driven by β-catenin flox/+ /LysM Cre/+ (β-cateninΔ/+) mice induces hypomyelination. In ( e ) quantification of Mbp mRNA by RT-qPCR at P10 in the anterior brain of β-catenin Δ/+ mice. mRNA levels arepresented as a fold change relative to WT mice, mean±SEM, Mann–Whitney test: * p<0.05. In ( f ) theβ-catenin deficit in β-catenin Δ/+ mice itself induces a decrease of myelin-binding protein MBP immunoreactivity (MBP-IR) in the corpus callosum and cingulum at P30 relative to Ctrl (β-catenin Δ/+ /C57Bla6) mice. Results are expressed by grey density, mean±SEM, Mann–Whitney test: *p<0.05. In ( g ) representative images of MBP-IR in the corpus callosum and cingulum at P30 in Ctrl and β-catenin Δ/+ . Scale bar: 60μm. For all bar graphs the n for each data point is indicated on or above the bar.

Article Snippet: To inhibit the Wnt/β-catenin pathway, we used XAV939 (Sigma-Aldrich, France).

Techniques: In Vivo, Gene Knockout, Injection, Isolation, Activation Assay, Quantitative RT-PCR, MANN-WHITNEY, Binding Assay

Journal: Cell Stem Cell

Article Title: Commitment and oncogene-induced plasticity of human stem cell-derived pancreatic acinar and ductal organoids

doi: 10.1016/j.stem.2021.03.022

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Article Snippet: XAV939 , Tocris , 3748.

Techniques: Activity Assay, Colorimetric Assay, Staining, Plasmid Preparation, Recombinant, Software, Cell Recovery

Journal: EMBO Reports

Article Title: Transposable element activity captures human pluripotent cell states

doi: 10.1038/s44319-024-00343-y

Figure Lengend Snippet: Reagents and tools table

Article Snippet: XAV939 , Axon Medchem , 1527.

Techniques: Recombinant, Modification, Purification, Sequencing, Knock-Out, Software, Electroporation