Journal: Frontiers in Molecular Neuroscience

Article Title: Major Signaling Pathways in Migrating Neuroblasts

doi: 10.3389/neuro.02.007.2009

Figure Lengend Snippet: In vitro analysis of the cytoskeleton pathway . (A) Cytoskeleton pathway involved in neuroblast migration resulting from the microarray data analysis. (B) Boyden chamber migrational assay: dissected neuroblasts were plated on a porous membrane, were allowed to migrate for 24 hours and Tuj1-positive cells were then counted ( p < 0.001). (C,D) Migration analysis in organotypic cultures obtained from 5HT3 A -EGFP mice. (C1) PI3K inhibitor (LY294002) severely disturbed neuroblast migration. Migration was quantified as the ratio of the EGFP-positive neuroblast containing area surrounding the SVZ between untreated (control) and treated (inhibitor) slices (obtained from the same sagittal level) after 4 days in culture ( n ≥ 5 slices per condition). (C2) Higher magnifications of RMS and SVZ in control and PI3K inhibitor-treated slices. Insets: note that LY294002-treated neuroblasts have short or no neurite compared to control neuroblasts. (C3) Quantification of the LY294002 effect ( n = 5, p < 0.001). (D1) Effect of PKCζ inhibitor. (D2) Directionality of neuroblast migration and, therefore, cell polarization is disturbed after PKCζ inhibitor treatment. Neuroblasts do not migrate in the streams, but migrate in all directions. This is clearly visible in the cortex where there are significantly more neuroblasts in PKCζ inhibitor-treated slices compared to control slices. (D3) Quantification of the PKCζ inhibitor effect ( n = 6, p < 0.005) Abbreviations: Ak – Akt1 inhibitor, C – control, cx – cortex, hp – hippocampus, lv – lateral ventricle, LY – PI3K inhibitor LY294002, P3,4,5 – PIP3,4,5; P4,5 – PIP4,5; P3,4 – PIP3,4; PZ – PKCζ inhibitor, Ra – Rac1 inhibitor, TA – Rho GTPases inhibitor Toxin A of C. difficile , W – PI3K inhibitor wortmannin.

Article Snippet: The following antibodies were used in our analysis: polyclonal rabbit anti-EGFP antibody, 1:10000 (Molecular Probes, USA), mouse anti-III class β-tubulin, Tuj1, 1:500 (Covance, USA), goat anti-CaM I, 1:200 (Santa Cruz, Germany), goat anti-doublecortin, 1:500 (Santa Cruz, Germany), rabbit anti-Akt1, 1:200 (Cell Signaling, USA), mouse anti-Wave1, 1:1000 (Neuromab, USA), rabbit anti-Cdc42, 1:1000 (Santa Cruz, Germany), rabbit anti-PI3K, 1:2000 (Upstate, USA), mouse anti-Rac1 (Cytoskeleton, USA), Alexa 488-conjugated anti-rabbit and anti-mouse secondary antibodies (Molecular Probes, USA), anti-mouse, anti-rabbit and anti-goat Cy3 coupled secondary antibodies (Jackson Immuno Research Laboratories, USA), anti-mouse and anti-rabbit HRP-conjugated secondary antibodies (Vector, USA).

Techniques: In Vitro, Migration, Microarray

Journal: Frontiers in Molecular Neuroscience

Article Title: Major Signaling Pathways in Migrating Neuroblasts

doi: 10.3389/neuro.02.007.2009

Figure Lengend Snippet: In vivo analysis of the cytoskeleton pathway . (A) Position of injection site (arrow) and destination area of migratory fluorescent cells (oval) after 7 or 10 days post-injection. (B) Western blot analysis of transfected HEK cells illustrating successful knockdown of Wave1 , Rac1 , Pik3r1 and Akt1 . (C) Red fluorescent cells in olfactory bulb infected by shRNAScrambled and shRNAAkt1 viruses. Fewer infected cells were found in OB of shRNAAkt1-injected animals. (D) Percentage of infected cells in olfactory bulb relative to total number of infected cells on SVZ-RMS-OB route after injection of shRNA expressing viruses against genes of the cytoskeleton pathway (* p < 0.005). Gene names are under histogram. GCL – granule cell layer, GL – glomerular layer.

Article Snippet: The following antibodies were used in our analysis: polyclonal rabbit anti-EGFP antibody, 1:10000 (Molecular Probes, USA), mouse anti-III class β-tubulin, Tuj1, 1:500 (Covance, USA), goat anti-CaM I, 1:200 (Santa Cruz, Germany), goat anti-doublecortin, 1:500 (Santa Cruz, Germany), rabbit anti-Akt1, 1:200 (Cell Signaling, USA), mouse anti-Wave1, 1:1000 (Neuromab, USA), rabbit anti-Cdc42, 1:1000 (Santa Cruz, Germany), rabbit anti-PI3K, 1:2000 (Upstate, USA), mouse anti-Rac1 (Cytoskeleton, USA), Alexa 488-conjugated anti-rabbit and anti-mouse secondary antibodies (Molecular Probes, USA), anti-mouse, anti-rabbit and anti-goat Cy3 coupled secondary antibodies (Jackson Immuno Research Laboratories, USA), anti-mouse and anti-rabbit HRP-conjugated secondary antibodies (Vector, USA).

Techniques: In Vivo, Injection, Western Blot, Transfection, Infection, shRNA, Expressing

Journal: Frontiers in Molecular Neuroscience

Article Title: Major Signaling Pathways in Migrating Neuroblasts

doi: 10.3389/neuro.02.007.2009

Figure Lengend Snippet: Phenotypes of animals infected into aSVZ/pRMS by AAV viruses expressing gene-specific shRNA and red fluorescent protein .

Article Snippet: The following antibodies were used in our analysis: polyclonal rabbit anti-EGFP antibody, 1:10000 (Molecular Probes, USA), mouse anti-III class β-tubulin, Tuj1, 1:500 (Covance, USA), goat anti-CaM I, 1:200 (Santa Cruz, Germany), goat anti-doublecortin, 1:500 (Santa Cruz, Germany), rabbit anti-Akt1, 1:200 (Cell Signaling, USA), mouse anti-Wave1, 1:1000 (Neuromab, USA), rabbit anti-Cdc42, 1:1000 (Santa Cruz, Germany), rabbit anti-PI3K, 1:2000 (Upstate, USA), mouse anti-Rac1 (Cytoskeleton, USA), Alexa 488-conjugated anti-rabbit and anti-mouse secondary antibodies (Molecular Probes, USA), anti-mouse, anti-rabbit and anti-goat Cy3 coupled secondary antibodies (Jackson Immuno Research Laboratories, USA), anti-mouse and anti-rabbit HRP-conjugated secondary antibodies (Vector, USA).

Techniques: Infection, Expressing, shRNA

Journal: iScience

Article Title: Tmem30a protects against podocyte injury through suppression of pyroptosis

doi: 10.1016/j.isci.2024.109976

Figure Lengend Snippet:

Article Snippet: RNase-free water , TIANGEN , Cat# RT121-02.

Techniques: Recombinant, Saline, Lysis, Western Blot, Marker, Membrane, Immunohistochemistry, Protein Concentration, Staining, Modification, Microarray, Software

Journal: Cell Reports

Article Title: Cytokine responsive networks in human colonic epithelial organoids unveil a molecular classification of inflammatory bowel disease

doi: 10.1016/j.celrep.2022.111439

Figure Lengend Snippet:

Article Snippet: RNase-Free DNase Set , Qiagen , 79254.

Techniques: Microarray, Recombinant, Cell Recovery, Sequencing, Software