cadherin 8 cdh8  (R&D Systems)

Journal: Biochemistry and Biophysics Reports

Article Title: Potential role of TGFΒ and autophagy in early crebellum development

doi: 10.1016/j.bbrep.2022.101358

Figure Lengend Snippet: Primary antibodies used for Immunoblotting.

Article Snippet: Anti- CDH8 , 1:1000 , Developmental Studies Hybridoma Bank Catalog# CAD8-1.

Techniques: Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Potential role of TGFΒ and autophagy in early crebellum development

doi: 10.1016/j.bbrep.2022.101358

Figure Lengend Snippet: Primer Sequences used for mRNA quantification.

Article Snippet: Anti- CDH8 , 1:1000 , Developmental Studies Hybridoma Bank Catalog# CAD8-1.

Techniques: Sequencing

Journal: Biochemistry and Biophysics Reports

Article Title: Potential role of TGFΒ and autophagy in early crebellum development

doi: 10.1016/j.bbrep.2022.101358

Figure Lengend Snippet: Cadherins expression are upregulated in cerebellar primordium during early developmental stages at E9-E13. A-H ) Expression levels of Cdh2, Ctnnb1, Cdh8 and Ncam1 proteins measured by Western blotting in mice cerebellar primordium at E9-E13 (n = 3). The protein loading was confirmed using β-actin. A, B ) Anti - Cdh2 reactive bands in Western blot analyses and quantification of Cdh2; Protein expression is upregulated from E9 to E11 and reached its higher amount at E11, which is statistically significant. C, D) Western blot analyses and quantification of Ctnnb1; protein expression remained unchanged with no significant differences between embryonic days. E, F ) Cdh8; protein blots show no expression during the earliest embryonic days at E9-E11. Cdh8 protein expression is upregulated from E12 to E13 and reached its higher amount at E13, which is statistically significant. G, H ) Data show no Ncam1 expression during the earliest embryonic days at E9-E11. Ncam1 protein expression is upregulated from E12 to E13 and reached its higher amount at E13 which is statistically significant. I-L ) mRNA expression levels of Cdh2 , Ctnnb1 , Cdh8 and Ncam1 measured by RT-qPCR in cerebellar primordium during early developmental stages at E9-E13 (n = 3). I ) Cdh2 mRNA expression remained unchanged with constant level of expression during the earliest embryonic days at E9-E13. J ) The trend for Ctnnb1 mRNA values is negative; transcription at E9 is higher with significant reduction at E11 and E12. K ) Cdh8 mRNA expression level experienced an upward trend from E9 to E13, with lower expression levels at E9/E10, which is significantly increased at E13. L ) Ncam1 mRNA levels increased from E9 to E13 with lower and higher levels of expression at E9 and E13, respectively. The data in the bar graphs are presented as the mean ± SEM, and statistical analysis was performed using one-way ANOVA (P-value ≤ 0.05 and lower were considered statistically significant. * shows P < 0.05, ** shows P < 0.01, *** shows P < 0.001, **** shows P < 0.0001). M-P ) Distribution of the Cdh2 , Ctnnb1 , Cdh8 and Ncam1 in the cerebellar primordium at E11.5, and E13.5. All images show cerebellar primordia and image credit: Allen Institute. © 2008 Allen Institute for Brain Science, Allen Developing Mouse Brain Atlas. Available online at: https://developingmouse.brain-map.org/ . M ) Cdh2 ISH data shows constant expression of Cdh2 mRNA at E11.5, and E13.5, with condense pattern in the nucmear transitory zone and Purkinje cell plate (mostly in Foxp2 + cells) at E13.5. N ) ISH data shows enrich expression of Ctnnb1 in the entire cerebellar primordium at E11.5 which is localized in the Purkinje cell plate at E13.5 (less in nuclear transitory zone). O ) Cdh8 mRNA is not detected at E11.5 but is detectable in caudal part of the Purkinje cell plate at E13.5. P ) Ncam1 ISH data shows strong expression pattern in the nuclear transitory zone of the cerebellar primordium at E11.5, and both nuclear transitory zone and Purkinje cell plate at E13.5.

Article Snippet: Anti- CDH8 , 1:1000 , Developmental Studies Hybridoma Bank Catalog# CAD8-1.

Techniques: Expressing, Western Blot, Quantitative RT-PCR

Journal: Biochemistry and Biophysics Reports

Article Title: Potential role of TGFΒ and autophagy in early crebellum development

doi: 10.1016/j.bbrep.2022.101358

Figure Lengend Snippet: The regulatory roles of Tgfb1 in formation of the cerebellar primordium during early developmental stages at E9-E13. Based on the neurogenesis timing of the cerebellum, the only postmitotic neural populations that exist in the cerebellar primordium between E9 to E13 are the Purkinje cells and cerebellar nuclei neurons located at the Purkinje cell plate and nuclear transitory zone, respectively. Expression of active Tgfb1 is intensely high at earliest embryonic days at both E9 and E10, followed by a substantial reduction in its value from E11 to E13. Our study shows that Tgfb1 through the canonical TGF-β signaling pathway could upregulate the expression of CAMs. On the other hand, activation of the Tgf-β signaling pathway and its consequent reduction in available Tgfb1 amounts, autophagic-flux, which the Tgf-β1 regularly induces, is inhibited from E9 to E13. A ) Activation of the Smad-dependent Tgf-β1 signaling pathway in the mouse cerebellar primordium at E9-E13; Our finding showed the presence of main components of the canonical Tgf-β signaling pathway, including Tgfbr1, Tgfbr2and cytoplasmic total and phosphorylated Smad signaling molecules during the given embryonic period. B ) Tgfb1 upregulates the expression of Cadherins in the mouse cerebellar primordium at E9-E13; Tgfb1 by upregulating the expression of cell adhesion proteins, including cdh2, Cdh8, Ncam, and cadherin binding protein β-catenin (Ctnnb1), contribute to the physical interactions and connectivity between postmitotic Purkinje cells and cerebellar nuclei neurons, locating in the Purkinje cell plate and nuclear transitory zone during earliest developmental stages from E9 to E13 with a peak at E11. These data show the direct association between upregulation of both Cdh2 and Ctnnb1, Cdh8 and Ncam upon activation of the Tgf-β signaling pathway, correlated to time points during which Purkinje cells and cerebellar nuclei neurons are born, migrating and positioning in cerebellar primordium. C ) Tgfb1 inhibits autophagic-flux in the mouse cerebellar primordium at E9-E13; Tgfb1, by regulating the autophagic-flux, responds to the extracellular signals such as stress (starvation, hypoxia, aggregation of unwanted materials) to maintain cellular homeostasis and provide neural protection during cerebellar development. The results from our study pinpoint a potential regulatory role of autophagy inhibition by Smad-dependent Tgf-β signaling pathway in the formation of the Purkinje cell plate and accumulation of cerebellar nuclei neurons in nuclear transitory zone during early cerebellar development.

Article Snippet: Anti- CDH8 , 1:1000 , Developmental Studies Hybridoma Bank Catalog# CAD8-1.

Techniques: Expressing, Activation Assay, Binding Assay, Inhibition

Journal: Journal of visualized experiments : JoVE

Article Title: Using microarrays to interrogate microenvironmental impact on cellular phenotypes in cancer.

doi: 10.3791/58957

Figure Lengend Snippet: The full list of ECM proteins and conditions that are used in the MEMA experiments. The uniprot ID, stock concentrations, and final working concentrations are provided. In some instances, the printed condition represents a protein complex or a combination of multiple proteins, which is indicated in the notes column.

Article Snippet: Cadherin-8 (CDH8) , R & D Systems , 188-C8 , ECM.

Techniques: Concentration Assay

Journal: Journal of visualized experiments : JoVE

Article Title: Using microarrays to interrogate microenvironmental impact on cellular phenotypes in cancer.

doi: 10.3791/58957

Figure Lengend Snippet: Materials

Article Snippet: Cadherin-8 (CDH8) , R & D Systems , 188-C8 , ECM.

Techniques: Imaging, Inverted Microscopy, Microarray, Electron Microscopy