Journal: Neurotherapeutics
Article Title: Regulatory T cells attenuate astrogliosis via IL-10/STAT3/PKC/vimentin signaling and promote neurological recovery after spinal cord injury
doi: 10.1016/j.neurot.2025.e00827
Figure Lengend Snippet: IL-10 supplementation and PKC phosphorylation inhibition recapitulate Treg-mediated downregulation of vimentin in vitro . A Representative immunofluorescence images of Foxp3 (green) and IL-10 (red) of magnetically sorted Tregs from mouse spleen. B ELISA quantification of IL-10 levels in supernatants of isolated Tregs. Cohen's d = 4.7545. C Experimental timeline of in vitro IL-10 intervention in astrocytes (rIL-10: recombinant IL-10; αIL-10, IL-10 neutralizing antibody). D-H Western blot quantification of p-STAT3/STAT3 (η 2 = 0.8939), SOCS3 (η 2 = 0.9199), p -PKC/PKC (η 2 = 0.9414), and vimentin (η 2 = 0.9252) protein levels in astrocytes following IL-10 intervention. I Representative immunofluorescence images of vimentin in astrocytes following IL-10 intervention. J Experimental timeline of in vitro PKC phosphorylation modulation in astrocytes (Go6976: PKC inhibitor; PMA: PKC activator). K, L Western blot quantification of vimentin protein levels in astrocytes following PKC modulation. η 2 = 0.8597. M, N Immunofluorescence quantification of vimentin levels in astrocytes following PKC modulation. η 2 = 0.6114. Data are mean ± SEM (n = 6 independent cultures). Statistical analyses were performed using two-tailed Student's t -test (B) or one-way ANOVA (E-H, L, N). P values are indicated on the graphs. n.s., not significant.
Article Snippet: For in vitro PKC modulation, we established optimal concentrations through gradient testing: astrocytes were treated with the PKC inhibitor Go6976 (100 nM, Selleck) for 24 h or the PKC activator Phorbol 12-myristate 13-acetate (PMA; 10 nM, Selleck) for 30 min.
Techniques: Phospho-proteomics, Inhibition, In Vitro, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Isolation, Recombinant, Western Blot, Two Tailed Test