Journal: Bioactive Materials

Article Title: Bioengineered extracellular vesicles escape lysosomal degradation and deliver Tet-PKM2 for macrophage immunometabolic reprogramming and periodontitis treatment

doi: 10.1016/j.bioactmat.2026.01.002

Figure Lengend Snippet: Generation of PKM2-overexpressing cells and generation of Tet-PKM2-enriched LEVs. ( A ) Schematic illustration depicting the overexpression of PKM2 in HEK293T cells by lentiviral transduction and subsequent allosteric activation of Tet-PKM2 via TEPP-46 treatment. Cells transfected with ov-NC or ov-PKM2 were named the NC or OV, respectively. ( B ) Relative mRNA expression levels of PKM2 in the NC and OV groups (qRT‒PCR, n = 4). ( C ) Representative immunoblot bands of PKM2 in the NC and OV groups. ( D ) Semiquantitative analysis of the expression levels of PKM2 in the NC and OV groups ( n = 6). ( E ) PK activity in PKM2-overexpressing HEK293T cells in response to treatment with various concentrations of TEPP-46 (0, 10, 20, 40, and 70 μM) ( n = 6). ( F ) Representative immunoblot bands of PKM2 conformational states in PKM2-overexpressing cells after treatment with TEPP-46 (40 μM) for 0, 8, and 24 h (DSS cross-linking assay). ( G ) Semiquantitative analysis of the expression levels of Tet-PKM2 in PKM2-overexpressing cells after treatment with TEPP-46 (40 μM) for 0, 8, and 24 h ( n = 4). ( H ) Schematic illustration of the isolation of LEVs and SEVs by differential velocity centrifugation. ( I ) Representative TEM images showing the morphology of SEVs and LEVs. ( J ) Size distributions of SEVs and LEVs (NTA). ( K ) Particle counts of SEVs and LEVs (NTA). ( L ) Particle-to-protein ratios of SEVs and LEVs. ( M ) Cellular uptake of SEVs and LEVs (labeled with PKH67; green) by macrophages (immunofluorescence assay); cell skeletons were stained with phalloidin (red), and nuclei were stained with DAPI (blue). ( N ) Expression of CD63, HSP70, TSG101, calnexin, and GM130 in whole-cell lysates (Cells), SEVs, and LEVs (Western blot). ( O ) Representative immunoblot bands of PKM2 conformational states in LEVs and SEVs. ( P ) Semiquantitative analysis of the expression levels of Tet-PKM2 in LEVs and SEVs ( n = 3) ( Q ) Conformational states of PKM2 in LEVs Tet−PKM2 in response to TEPP-46 treatment. ( R ) Semiquantitative analysis of the expression levels of Tet-PKM2 in LEVs following TEPP-46 treatment ( n = 3). The data are expressed as the mean ± SEM. Statistical analysis was performed by one-way ANOVA ( E and G ) and Student's t -test ( B , D , K , L , P , and R ). ns indicates no significant difference between the indicated columns; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 indicate significant differences between the indicated columns.

Article Snippet: PKH67-labeled SEVs and LEVs resuspended in complete medium were used to treat RAW 264.7 cells for 24 h. The cells were then fixed with 4 % PFA (Coolaber), permeabilized, and stained for cytoskeletal visualization using fluorescein phalloidin (1:1000 dilution; MCE) for 30 min.

Techniques: Over Expression, Transduction, Activation Assay, Transfection, Expressing, Western Blot, Activity Assay, Isolation, Centrifugation, Labeling, Immunofluorescence, Staining

Journal: Genes & Diseases

Article Title: cGAS knockout inhibited endotoxin-induced uveitis in mice

doi: 10.1016/j.gendis.2025.101786

Figure Lengend Snippet: Representative fundus photographs (A–D) and FFA (E–H) of the retinas in the NC and EIU groups. (A) Fundus photograph from the WT NC group. (B) Fundus photograph from the KO NC group. (C) Fundus photograph from the WT EIU group. (D) Fundus photograph from the KO EIU group. (E) FFA from the WT NC group. (F) FFA from the KO NC group. (G) FFA from the WT EIU group. (H) FFA from the KO EIU group. (I) Grade of the severity of vitreous opacity. ∗∗ P < 0.01. n = 3 biological replicates per group. FFA, fundus fluorescein angiography; WT, wild type; KO, Cgas knockout; NC, normal control; EIU, endotoxin-induced uveitis.

Article Snippet: Concanavalin A and fluorescein (FL-1001-25; Vectorlabs) were dissolved in PBS.

Techniques: Knock-Out, Control