Journal: The FASEB Journal

Article Title: CSPG4 Mediates Inflammatory, Cell Death, and Senescence Responses in Enteric Glia Exposed to Clostridioides difficile Toxins

doi: 10.1096/fj.202600333R

Figure Lengend Snippet: CSPG4 blockade reduces TcdB‐induced NFκB nuclear translocation and IL‐6 gene expression. (A) Representative photomicrographs of NFκB (green) immunostaining, phalloidin (red) as a cytoplasm staining, and DAPI (blue) as a nuclear staining in enteric glia cells exposed to TcdA (50 ng/mL) or TcdB (1 ng/mL) at 18 h incubation in the presence of isotype control or anti‐CSPG4 antibodies (0.001 μg/mL). Scale bars: 50 μm. (B) Percentage of enteric glia cells ( n = 5, mean ± SEM) with positive nuclear NFκBp65 staining at 18 h of incubation with TcdA and TcdB in the presence of isotype control or anti‐CSPG4 antibodies. (C) IL‐6 gene expression (mean ± SEM) by qPCR in enteric glial cells challenged with TcdA or TcdB at 18 h of incubation in the presence of isotype control or anti‐CSPG4 antibodies (0.001 μg/mL). (B‐C) One‐way ANOVA followed by the Tukey test.

Article Snippet: Slides were mounted using antifade mounting medium containing phalloidin (Vector Labs, H1600) to visualize the actin cytoskeleton.

Techniques: Translocation Assay, Gene Expression, Immunostaining, Staining, Incubation, Control

Journal: The FASEB Journal

Article Title: CSPG4 Mediates Inflammatory, Cell Death, and Senescence Responses in Enteric Glia Exposed to Clostridioides difficile Toxins

doi: 10.1096/fj.202600333R

Figure Lengend Snippet: CSPG4 blockade reduces TcdB‐induced pSTAT3 nuclear translocation in enteric glial cells. (A) Representative photomicrographs of pSTAT3 (green) immunostaining, phalloidin (red) as a cytoplasm staining, and DAPI (blue) as a nuclear staining in enteric glial cells exposed to TcdA (50 ng/mL) or TcdB (1 ng/mL) at 18 h incubation in the presence of isotype control or anti‐CSPG4 antibodies (0.001 μg/mL). Scale bars: 50 μm. (B) Percentage of enteric glial cells ( n = 5, mean ± SEM) with positive nuclear pSTAT3 staining at 18 h of incubation with TcdA and TcdB in the presence of isotype control or anti‐CSPG4 antibodies (0.001 μg/mL). One‐way ANOVA followed by the Tukey test.

Article Snippet: Slides were mounted using antifade mounting medium containing phalloidin (Vector Labs, H1600) to visualize the actin cytoskeleton.

Techniques: Translocation Assay, Immunostaining, Staining, Incubation, Control

Journal: STAR Protocols

Article Title: Combining electrophysiology and optogenetics for functional screening of pyramidal neurons in the mouse prefrontal cortex

doi: 10.1016/j.xpro.2021.100469

Figure Lengend Snippet: Experimental setup and essential equipment for in utero electroporation (A) Image of the whole setup for in utero electroporation. (B) Images of the electroporator to deliver electrical stimulation to pups (upper) and the micro injector to inject the plasmids (lower). (C) Images of surgical implements and a glass pipette which contains the diluted solution including plasmids for in utero electroporation. (D) Scheme of the in utero electroporation. The plasmid is injected into the fetal lateral ventricle and electrical stimulation is delivered via the forceps-shaped bipolar electrode. The electroporation is performed on 6–8 embryos in a single operation. (E) Inject the solution containing the plasmid into one side of the lateral ventricles on each embryo. The injection site of the plasmid should be 0.75–1.0 mm anterior from lambda and 0.5 mm lateral from the sagittal suture. (F) The positions of the electrode to target the mPFC. Note that the negative pole is positioned over the injected hemisphere while the positive one is positioned on the contralateral side. The two electrodes are slightly angled rostrocaudally and vertically as seen. (G–I) Low (G) and high (H) magnification Images of injection of the plasmids and image of delivering electrical stimulation through a forceps-shape electrode (I) during in utero electroporation. (J) Image of the mPFC from an electroporated mouse. The green signal shows GFP and the blue signal represents DAPI staining. Scale, 1mm. (K) High-magnification image of the electroporated mPFC expressing GFP. Scale, 50 μm. See also .

Article Snippet: VECTASHIELD® HardSet TM Antifade Mounting Medium with DAPI , Merck Millipore , CAT: # H-1500-10.

Techniques: In Utero, Electroporation, Transferring, Plasmid Preparation, Injection, Staining, Expressing

Journal: STAR Protocols

Article Title: Combining electrophysiology and optogenetics for functional screening of pyramidal neurons in the mouse prefrontal cortex

doi: 10.1016/j.xpro.2021.100469

Figure Lengend Snippet:

Article Snippet: VECTASHIELD® HardSet TM Antifade Mounting Medium with DAPI , Merck Millipore , CAT: # H-1500-10.

Techniques: Recombinant, Mutagenesis, Transfection, Blocking Assay, Expressing, Plasmid Preparation, DNA Purification, cDNA Synthesis, Ligation, Software, In Utero, Electroporation, Microscopy