sema3a pathway components sema3a  (Shanghai Korain Biotech Co Ltd)

Journal: Neural Regeneration Research

Article Title: Activin A enhances neurofunctional recovery following traumatic spinal cord injury by inhibiting autophagy

doi: 10.4103/NRR.NRR-D-24-01021

Figure Lengend Snippet: AA alleviates excessive autophagy via activation of the PI3K pathway. (A, B) Intracellular deposition of the autophagy marker LC3-II captured by imaging flow cytometry. Increased LC3-II deposition was observed in the cytosol of H 2 O 2 -treated cells, which was alleviated by AA treatment. Bar plots show each group’s relative fluorescence intensity and LC3-II deposition area ( n = 6). (C) Western blot images show the expression level of autophagy-related proteins in cell samples. (D) Quantitative analysis of the expression content of autophagy-related proteins in C. (E) Representative immunofluorescence images of LC3-II (green, Alexa Fluor® 488) deposition in spinal cord sections. The injury center in the AA treatment group had a smaller LC3-II deposition area than did controls. (F) Representative western blot images of autophagy markers (LC3 I/II, and P62) and a semaphorin family member (Sema3A) in spinal cord tissue. (G) Quantitative analysis of fluorescence intensity in E and WB bands in F. (H) qPCR showing the expression level of the Sema3A and receptor protein Nrp1 ( n = 3). (I) Representative immunofluorescence images of Sema3A + (green, Alexa Fluor® 488) cells after H 2 O 2 stimulation and AA treatment. Sema3A was highly expressed in neuronal cytosol and axons. Cells that received AA treatment had higher Sema3A expression compared with those in the H 2 O 2 group, and the increase was concentration-dependent. (J) Representative immunofluorescence images of Nrp1 (red, Alexa Fluor® 555) and Sema3A (green, Alexa Fluor® 488) in spinal cord sections. The AA-treated group exhibited more pronounced Sema3A/Nrp1-positive signals at the injury site and a more intact tissue junction than did the SCI group. (K) Western blot images showing the expression levels of PI3K/AKT/mTOR pathway-related proteins. (L) Quantitative analysis of the expression levels of PI3K/AKT pathway-related proteins in K. (M) The expression levels of EGFR/PI3K pathway-related genes by qPCR ( n = 3). Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance with Tukey’s multiple comparisons test for B, D, and L; one-way analysis of variance with Dunnett’s multiple comparisons test for H and M). Scale bars: as shown in the Figure. AA: Activin A; AKT: protein kinase B; BF: bright field; DAPI: 4′,6-diamidino-2-phenylindole dihydrochloride; EGFR: epidermal growth factor receptor; LC3: microtubule-associated proteins light chain; mTOR: mammalian target of rapamycin; Nrp1: neuropilin 1; p-AKT: phospho-AKT; p-mTOR: phospho-mTOR; p-PI3K: phospho-PI3K; PBS: phosphate-buffered saline; PI3K: phosphoinositide-3-kinase; pik3cg: PI3K catalytic subunit gamma; qPCR: quantitative polymerase chain reaction; SCI: spinal cord injury; Sema: semaphorin; SP1: specificity protein 1; WB: western blot.

Article Snippet: Sema3A , Mouse , 1:500 , ABclonal, Woburn, MA, USA , A12967 , AB_2759814 , WB, IF.

Techniques: Activation Assay, Marker, Imaging, Flow Cytometry, Fluorescence, Western Blot, Expressing, Immunofluorescence, Concentration Assay, Saline, Real-time Polymerase Chain Reaction

Journal: eLife

Article Title: Human receptive endometrial assembloid for deciphering the implantation window

doi: 10.7554/eLife.90729

Figure Lengend Snippet: ( A ) GSEA between the secretory (SEC) and window of implantation (WOI) groups for proliferative epithelium. ( B ) The single-cell pseudotime trajectory of proliferative epithelium, stromal cell, EMT-derived cell and stem-derived epithelium. Cells start at proliferative epithelium and progress to EMT-derived cell. There are seven major states over pseudotime. The black spot indicates the differentiation node between states 4, 5 and 6, indicating the direction from proliferative epithelium to EMT-derived cell. Arrows indicate the direction of the pseudotime trajectory. ( C ) The horizontal axis is the pseudotime point, and the vertical axis is the gene expression level. The solid line represents states 1, 2, 4, and 5 corresponding to Fig. S4B. Different colors represent samples in the CTRL, SEC, and WOI groups. ( D ) Heatmap of genes at the branch node regulating differentiation into EMT-derived cell and stem-derived epithelium. The horizontal axis is the pseudo-time point (the pseudo-time point gradually increases from the middle to both sides). The vertical axis is the gene expression level, representing two differential directions on the left and right sides. Clusters represent the gene sets with a similar branch gene expression trend. Different colors represent the level of gene expression. ( E–F ) Dot plots demonstrating the Cellphone DB analysis of relevant receptors and ligands of EMT-derived cell ( E ) or stromal cell ( F ) with other cell types. The size of the dot represents the level of significance. The color of the dot indicates the mean of the average expression level of interacting molecule 1 in EMT-derived cells ( E ) or stromal cells ( F ) and molecule 2 in other cell types. ( G ) Proximity ligation assay (PLA) validating the interactions of SEMA3A-NRP1 and CD46-JAG1 in the CTRL, SEC and WOI assembloids. Red signals the interaction of two proteins. Nuclei were counterstained with DAPI. Scale bar = 20 μm.

Article Snippet: Antibody , SEMA3A antibody (C-1) (Mouse monoclonal) , Santa Cruz , Cat#: sc-74555 RRID: AB_2185394 , PLA (1:50).

Techniques: Single Cell, Derivative Assay, Gene Expression, Expressing, Proximity Ligation Assay