Journal: Molecular Neurobiology

Article Title: Comparative Analysis of Red Onion-Derived Exosome-Like Nanovesicles and Extract Reveals Sustained Immunomodulatory Effects in LPS/IFN-γ-Stimulated Microglia

doi: 10.1007/s12035-026-05820-0

Figure Lengend Snippet: Anti-inflammatory and antioxidant properties of RO-ELNs and extract against ATP + LPS/IFN-γ stimulated pro-inflammatory mediators in HMC3 cells. RO-ELNs with a concentration of 1 × 10 9 particles/mL were pretreated for 20 or 6 h before acute or prolonged neuroinflammation models. Extract with a concentration of 125 µg/mL was incubated for only 6 h for both acute and prolonged neuroinflammation models. Western blot and quantitative analyses show the ability of RO-ELNs and extract in inhibiting P-NF-κB ( A , B ) and COX-2 ( C , D ) expression levels in HMC3 cells at different time points 4 and 48 h of neuroinflammation. GAPDH was used as a housekeeping protein. The antioxidative properties of RO-ELNs and extract ( E ) and their ability to decrease IL-6, TNF-α secretion levels ( F , G ) at 4 and 48 h of neuroinflammation. Data are expressed as mean ± standard ( n ≥ 3): * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. As a result of the initial reversed loading sequence for COX-2, lanes were rearranged digitally to align with the experimental grouping presented in the paper. All lanes originated from the same membrane and exposure. A separation dash marks the realignment. Uncropped original membranes can be found in Supplementary Figure

Article Snippet: 3033), and GAPDH (Boster, M00227-7) antibodies overnight at 4 °C.

Techniques: Concentration Assay, Incubation, Western Blot, Expressing, Sequencing, Membrane

Journal: Molecular Therapy. Nucleic Acids

Article Title: Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury

doi: 10.1016/j.omtn.2018.03.009

Figure Lengend Snippet: Downregulation of GSK3B and RhoA in DRG Explants DRG explants were cultured within a 3D fibrin hydrogel loaded with LNA-AONs (6 µM inside the gel or 2 µM if related to final culture volume: gel/20 µL + culture medium/40 µL). 6 μM inside the gel or 2 μM if related to final culture volume: gel/20 μL + culture medium/40 μL. (A) Relative RNA quantification by qRT-PCR after a 7-day exposure to LNA AONs. LNA AONs against an irrelevant sequence (LNA6424-GFP) were used as controls. Each point represents an independent experiment where RNA from a pool of 10–15 independently treated DRG explants was used per condition and per independent experiment for qPCR quantification. Results indicate mean ± SD. One way-ANOVA with Dunnett’s multiple comparison test (versus non-treated [NT]) was used where indicated (*p < 0.05; **p < 0.01; n.s., not significant). (B) Western blot analysis of GSK3β and RhoA protein expression levels after AON treatments. Protein was extracted from a pool of 10–15 DRG explants, each treated independently with LNA gapmers. Percentages shown indicate the relative amount of protein remaining in comparison to control and normalized to the GAPDH band, as calculated by semiquantitative analysis (band densitometry). For the original western blot membranes, see also Figure S2 .

Article Snippet: The primary antibodies and specific dilutions used were as follows: mouse anti-GSK3 α/β (1:2,000; Santa Cruz Biotechnology), rabbit anti-RhoA (1:1,000; Cell Signaling Technology), mouse anti-GAPDH (1:40,000; HyTec).

Techniques: Cell Culture, Quantitative RT-PCR, Sequencing, Western Blot, Expressing