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: After washing, cells were incubated overnight at 4°C with primary antibodies against CSPG4 (Abcam, ab275024, 1:400), nectin‐3 (PVRL3, Invitrogen, PA5‐51095, 1:200), LRP1 (Abcam, ab92544, 1:100), NFκB p65 (Santa Cruz Biotechnology, sc‐372, 1:100), or pSTAT3 (R&D Systems, AF4607, 1:100).

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