Journal: bioRxiv
Article Title: Evaluation of Tetraspanins in Extracellular Vesicle Bioengineering
doi: 10.64898/2026.01.13.699196
Figure Lengend Snippet: Lentiviral transduction to generate WT, PanKO, CD9KO, CD63KO, and CD81KO cells expressing TlucCD63-Cerulean. A. Schematic workflow of engineered Tluc-EVs by introducing TlucCD63-Cerulean lentiviruses (created with BioRender.com ). B. The percentage of Cerulean positive cells. C. The mean fluorescent intensity (MFI) of Cerulean positive cells. D. Flow cytometry plot for the cells after staining with APC-conjugated CD9/CD63/CD81 tetraspanin antibodies. E. Fold increase of engineered Tluc-CD63 EVs in PanKO-, CD9KO, CD63KO-, and CD81KO-cells over WT cells (Data are normalized to the RLU of EVs from WT cells). F. Fold increase of engineered Tluc-CD63EVs in between PanKO-, CD9KO, CD63KO-, and CD81KO-cells (Data are normalized to the RLU of EVs from WT cells and compared across KO groups). G. Heatmap of expressing tetraspanins in EVs. H. Interaction network of CD63 with the tetraspanins retrieved from STRING. The data are presented as means (±SD, n = 3-5). One-way ANOVA was used to show significance and was illustrated as follows: * p < 0.05, ** p < 0.01, **** p < 0.0001.
Article Snippet: Briefly, 2.5 × 10 8 EVs, quantified by NTA, were stained with 8 nM fluorescent antibodies targeting the tetraspanins CD9-APC (clone: SN4 C3-3A2, cat. 130-128-037), CD63-APC (clone: H5C6, cat. 130-100-182, Miltenyi Biotec), and CD81-APC (clone: JS64, Beckman Coulter).
Techniques: Transduction, Expressing, Flow Cytometry, Staining