Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: The exosomal protein NID1 promotes liver metastasis of CRC tumors (A) EV mouse model comparing the effects of EVs from SW480/km12-LM3 and shNID1-1 SW480/km12-LM3 cells on CRC metastasis ( n = 5). (B and C) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (D) Comparison of the effects of EVs from XPack and XP-NID1 HCT116 cells on CRC metastasis in an EV mouse model ( n = 5). (E and F) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (G) Schematic diagram of the mouse model of in situ cecum injection combined with splenic injection. (H and I) In vivo imaging images of the mice and quantitative fluorescence statistics of the mouse liver region. n.s. represents not significant; ∗∗ represents p < 0.01; ∗∗∗ represents p < 0.001, analyzed with a t test (B, C, E, F, H, and I). Data are represented as mean ± SEM.

Article Snippet: NID1 was expressed through XPack EV Protein Engineering Technology (System Biosciences) for expression in the EVs of HCT116 and NCM-460 cells.

Techniques: Luciferase, Comparison, In Situ, Injection, In Vivo Imaging, Fluorescence

Journal: Molecular Therapy

Article Title: Hypoxia-induced lncHILAR promotes renal cancer metastasis via ceRNA for the miR-613/206/ 1-1-3p/Jagged-1/Notch/CXCR4 signaling pathway

doi: 10.1016/j.ymthe.2021.05.020

Figure Lengend Snippet: Exosomes derived from hypoxic RCC cells carry lncHILAR and promote cell invasion by normoxic RCC cells (A) The cellular distribution of lncHILAR RNA by qRT-PCR showed that lncHILAR was predominately localized to the cytoplasm. (B) RNAscope analysis was used to explore the distribution of lncHILAR in RCC samples. (C) qRT-PCR analysis of lncHILAR in the cell medium of ACHN and Caki-1 cells treated with RNase (2 mg/mL) alone or combined with Triton X-100 (0.1%) for 20 min. n = 3. ∗∗∗p < 0.0001. (D) qRT-PCR analysis of lncHILAR in the exosomes of ACHN and Caki-1 cells cultured under normoxia and hypoxia. n = 3. ∗p < 0.05. (E) Exosome uptake assay was confirmed with XPack MSCV-XP-GFP-EF1α-Puro Expression LentiVector. GFP-labeled exosomes were taken up by ACHN cells, shown as the green signal. (F) Cells were incubated with CM for 24 h. qRT-PCR analysis of lncHILAR of ACHN and Caki-1 cells after treatment with different CMs. CM+GW4869, CM collected from cells pretreated with GW4869 (5 μM). n = 3. ∗p < 0.05. (G) For in vitro exosome treatment, exosomes collected from about 5 × 10 6 producer cells were added to 2 × 10 5 recipient cells in each plate of a six-well plate. The cells were then incubated with exosomes in CM for 24 h. qRT-PCR analysis of lncHILAR of ACHN and Caki-1 cells after treatment with hypoxic exosomes. NM, normal medium. n = 3. ∗∗∗p < 0.0001. (H) Cell invasion analysis of ACHN and Caki-1 cells treated with different CM. n = 3. ∗p < 0.05. (I) Transwell invasion assay of ACHN cells, Caki-1 cells, ACHN lncHILAR-KD cells, and Caki-1 lncHILAR-KD cells after treatment with hypoxic exosomes. n = 3. Caki-1 Exo, hypoxic exosomes derived from hypoxic Caki-1 cells; ACHN Exo, hypoxic exosomes derived from hypoxic ACHN cells. n = 3. ∗p < 0.05.

Article Snippet: ∗p < 0.05. (E) Exosome uptake assay was confirmed with XPack MSCV-XP-GFP-EF1α-Puro Expression LentiVector.

Techniques: Derivative Assay, Quantitative RT-PCR, RNAscope, Cell Culture, Expressing, Labeling, Incubation, In Vitro, Transwell Invasion Assay

Journal: Neuroglia

Article Title: Brain-Homing Peptide Expression on the Membrane Enhances the Delivery of Exosomes to Neural Cells and Tissue

doi: 10.3390/neuroglia6010003

Figure Lengend Snippet: Figure 3. The size profile analysis of exosomes captured based on CD9, CD63, and CD81 markers is shown as follows: (A) HEK293 (XPack GFP) exosomes processed using the PEG method, (B) HEK293 (XPack GFP) exosomes not processed using the PEG method, and (C) table displaying the mean and mode sizes of exosomes captured by CD9, CD63, and CD81 markers.

Article Snippet: The transfer plasmid containing the gene of interest, such as the XPack GFP (System Biosciences, Palo Alto, CA, USA) or XStamp BHP (System Biosciences, Palo Alto, CA, USA), was co-transfected with packaging plasmids (pLP1) and an envelope plasmid (VSV-G) into HEK 293TN cells using LipofectamineTM 2000 (ThermoFisher, Waltham, MA, USA) according to the manufacturer’s instructions.

Techniques:

Journal: Neuroglia

Article Title: Brain-Homing Peptide Expression on the Membrane Enhances the Delivery of Exosomes to Neural Cells and Tissue

doi: 10.3390/neuroglia6010003

Figure Lengend Snippet: Figure 4. (A) Flow cytometry analysis showing the percentage of GFP-positive cells following treat- ment with HEK 293 exosomes, HEK 293 exosomes (XPack GFP), and HEK 293 exosomes (XPack GFP + XStamp BHP). Fluorescent microscopy images of GBM CSCs post-exosome treatment: (B) control, (C) XPack GFP, and (D) XPack GFP + XStamp BHP. Dot blot analysis for GFP detection from brain lysates of NSG mice treated with HEK 293 (XStamp BHP) and HEK 293 (XPack GFP + XStamp BHP) exosomes. Statistical analysis was performed using a one-way ANOVA followed by Holm–Sidaks multiple comparison test. Significance levels were indicated as *** p < 0.001 and **** p < 0.0005.

Article Snippet: The transfer plasmid containing the gene of interest, such as the XPack GFP (System Biosciences, Palo Alto, CA, USA) or XStamp BHP (System Biosciences, Palo Alto, CA, USA), was co-transfected with packaging plasmids (pLP1) and an envelope plasmid (VSV-G) into HEK 293TN cells using LipofectamineTM 2000 (ThermoFisher, Waltham, MA, USA) according to the manufacturer’s instructions.

Techniques: Flow Cytometry, Microscopy, Control, Dot Blot, Comparison