Journal: Molecular Brain

Article Title: Ubiquilin-2 drives NF-κB activity and cytosolic TDP-43 aggregation in neuronal cells

doi: 10.1186/s13041-015-0162-6

Figure Lengend Snippet: hUBQLN2 co-localized with TDP-43 and P62 but not with NF-κB and IκB-α. Immunofluorescence of Neuro2A cells at 48 h after transfection with ( a ) control plasmid, ( b ) pCMV-hUBQLN2 WT and ( c ) pCMV-hUBQLN2 P497H . Indicated antibodies were used according to Materials and Methods. Microscopy pictures were taken at 63× magnification. d - e Western analysis from three repeated experiments was used for quantification of ( d ) TDP-43 level ( p = 0.0227 for hUBQLN2 WT and p = 0.0032 for hUBQLN2 P497H ) and ( e ) p62 level ( p = 0.0058 for hUBQLN2 WT and p = 0.0130 for hUBQLN2 P497H ) in insoluble fraction. Results were calculated in fold change compared to proteins level in insoluble fraction of control cells. f Neuro2A cells were transfected with control plasmid, pCMV-hUBQLN2 WT or pCMV-hUBQLN2 P497H . Cells were extracted 48 h after transfection using aggregates assay protocol. The results suggested that overexpression of UBQLN2 species increased levels of UBQLN2, TDP-43 and p62 in the insoluble fraction. g Live imaging was performed on Neuro2A pFeGFP-hUBQLN2 P497H and DsRED-TDP-43 co-transfected cells. Microscopy pictures were taken at 24 and 48 h after transfection. h Immunoblot of TDP-43 in cytoplasmic (C) vs nuclear (N) after transfection with control plasmid, pCMV-hUBQLN2 WT or pCMV-hUBQLN2 P497H plasmids. i HEK293 cells and Neuro2A cells were transfected with either control plasmid or pCMV-hUBQLN2 P497H plasmid. Protein levels of hUBQLN2 and mUBQLN2 were measured at 48 h after transfection. Human UBQLN2 levels were similar in control HEK293 and pCMV-hUBQLN2 P497H Neuro2A transfected cells. Scale bar = 25 μm

Article Snippet: Primary antibodies were monoclonal Flag M2 (Sigma-Aldrich, Saint-Louis, 1:100), TARDBP (Proteintech, Chicago, 1:600), NF-κB p65 (Santa-Cruz, 1:200), IκB-α (Santa-Cruz, 1:200) and SQSTM1/P62 (Cell signaling, 1:200).

Techniques: Immunofluorescence, Transfection, Control, Plasmid Preparation, Microscopy, Western Blot, Over Expression, Imaging

Journal: Journal of Neuroinflammation

Article Title: Roseburia intestinalis ameliorates adolescent depression via GPR43‑dependent Treg cell expansion and suppression of neuroinflammation

doi: 10.1186/s12974-026-03755-w

Figure Lengend Snippet: Schematic illustration of the experimental flow in vivo and in vitro experiments. In vivo study, a chronic restraint stress (CRS) mouse model was established and subjected to intervention with R.i. , followed by separate administration of an Free Fatty Acid Receptor 2 (FFAR2) inhibitor (GLPG0974) or a CD25-neutralizing antibody (Anti-mouse CD25 Antibody, PC61.5). Afterward, the inflammatory profiles, levels of short-chain fatty acids (SCFAs), immune and neural parameters in the hippocampus, cortex and colon, as well as intestinal metabolites were evaluated. In vitro experiment, the polarization of microglia was assessed in an LPS+corticosterone (LPS&CORT) induced cellular stress model followed by intervention with R.i. -conditioned medium

Article Snippet: GLPG0974 (HY-12940, MCE), a free fatty acid receptor 2 (FFAR2/GPR43) antagonist, was prepared as a 5 mg/mL working solution and administered intraperitoneally to mice at a dose of 100 μL every three days, for a total of 10 injections throughout the modeling period.

Techniques: In Vivo, In Vitro