cas  (Chem Impex International)

Journal: Journal of Extracellular Vesicles

Article Title: M1 Macrophage‐Derived Small Extracellular Vesicles as Synergistic Nanotherapeutics: Harnessing Intrinsic Anticancer Activity and Drug Delivery Capacity

doi: 10.1002/jev2.70242

Figure Lengend Snippet: Comprehensive sEV characterization and M1‐sEV Immune Evasion. (a) Fluorescence‐activated cell sorting (FACS) analysis of CD86 (M1 marker) and CD206 (M2 marker) in lipopolysaccharide (LPS; 500 ng/mL)‐stimulated M0 macrophages to confirm M1 polarization prior to sEV isolation. Transmission electron microscopy (TEM) images showing spherical nanostructures and corresponding size distribution analysis ( n = 80) of (b) 3T3‐sEVs, (c) M0‐sEVs, and (d) M1‐sEVs. (e) Western blot analysis of sEV markers and immunomodulatory ligand. Nanoparticle tracking analysis (NTA) showing size distribution and particle concentration profiles ( n = 5) of (f) 3T3‐sEVs, (g) M0‐sEVs, and (h) M1‐sEVs. Representative ex vivo fluorescence images and corresponding biodistribution quantification ( n = 3, mean ± s.d.) of major organs resected from mice administered with (i, m) Ctrl (PBS), (j, n) 3T3‐sEVs, (k, o) M0‐sEVs, and (l, p) M1‐sEVs. DiD‐labelled sEVs (measurement: ex/em = 605/670 nm). Quantification normalized to the fluorescence signal intensity of DiD‐3T3‐sEVs. One‐way ANOVA with Tukey's post‐test. ns: not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Fluorescence‐activated cell sorting system (FACS; Cytek Northern LightsTM (NL)‐CLC NL‐2000; Cytek Biosciences, CA, USA) was used to sort and detect the cells.

Techniques: Fluorescence, FACS, Marker, Isolation, Transmission Assay, Electron Microscopy, Western Blot, Concentration Assay, Ex Vivo

Journal: Journal of Extracellular Vesicles

Article Title: M1 Macrophage‐Derived Small Extracellular Vesicles as Synergistic Nanotherapeutics: Harnessing Intrinsic Anticancer Activity and Drug Delivery Capacity

doi: 10.1002/jev2.70242

Figure Lengend Snippet: Tumour‐homing properties of M1‐sEVs. (a) Mass spectrometry (MS)‐based proteomic profiling of sEVs. Protein accession numbers from UniProtKB/Swiss‐Prot; black boxes with [+]: detected proteins; white boxes with [−]: undetected protein; Gal‐3: Galectin‐3; VEGFR1: Vascular endothelial growth factor receptor‐1; NRP: Neuropilin; HSP: Heat shock protein; CHC: Clathrin heavy chain; CLC: Clathrin light chain. Fluorescence‐activated cell sorting (FACS) analysis of DiD‐labelled sEV uptake to MCF‐7 cells at (b) 1 h, (c) 4 h, and (d) 8 h at 37°C, with (f) corresponding quantification ( n = 3, mean ± s.d.). (e) FACS analysis of temperature‐dependent uptake at 4°C, with (g) corresponding quantification ( n = 3, mean ± s.d.). Quantification threshold: DiD fluorescence intensity > 10 4 . (h) Two‐photon confocal laser scanning microscopy (TP‐CLSM) images of DiD‐labeled sEV uptake in MCF‐7 spheroids in inverted colloidal crystal (iCC) framework (Blue: live cells; Green: 5‐DTAF‐labeled iCC; Red: DiD‐labelled sEV). (i) In vivo fluorescence imaging of DiD‐sEVs at various time points following tail‐vein intravenous (i.v.) administration in MCF‐7 tumour–bearing female NCr nude mice. (j) Quantitatively collected DiD‐sEV intensity in the spheroid penetration depth in TP‐CLSM images ( N > 30). (k) Time‐dependent quantification of DiD‐sEV accumulation in tumour based on in vivo systemic biodistribution imaging ( n = 4, mean ± s.d.). One‐way ANOVA with Tukey's post‐test. ns: not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Fluorescence‐activated cell sorting system (FACS; Cytek Northern LightsTM (NL)‐CLC NL‐2000; Cytek Biosciences, CA, USA) was used to sort and detect the cells.

Techniques: Mass Spectrometry, Fluorescence, FACS, Confocal Laser Scanning Microscopy, Labeling, In Vivo, Imaging

Journal: Cell death & disease

Article Title: Circular RNA TFRC/SCD1 mRNA interaction regulates ferroptosis and metastasis in gastric cancer.

doi: 10.1038/s41419-025-07759-x

Figure Lengend Snippet: Fig. 4 CircTFRC knockdown promoted the ferroptosis of GC cells. A, B Propidium iodide (PI) staining showing the cell death rates of control (si-NC) and circTFRC knockdown (si-circTFRC) AGS and HGC-27 cells in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). C, D Flow cytometry showing the lipid ROS levels (stained with C11 BODIPY 581/591) in control (si-NC) and circTFRC knockdown (si-circTFRC) AGS and HGC-27 cells in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). ROS reactive oxygen species. E, F ELISA assays showing the relative MDA levels in control (si-NC) and circTFRC knockdown (si-circTFRC) AGS and HGC-27 cells in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). MDA malondialdehyde. G, H Plate colony formation assay assessing colony formation in AGS cells under control conditions (sh-NC) or with circTFRC knockdown (sh-circTFRC) in the absence or presence of ferrostatin-1 (0.25 μM, 72 h). I, J Transwell assay evaluating the migration ability of AGS cells under control conditions (sh-NC) or with circTFRC knockdown (sh-circTFRC) in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). K PI staining showing the cell death rates of control (sh-NC) and circTFRC knockdown (sh-circTFRC) AGS cells following treatment with RSL3 (1 μM) in the absence or presence of ferrostatin-1 (0.75 μM) for 16 h. L Flow cytometry showing the lipid ROS levels (stained with C11 BODIPY 581/591) in control (sh-NC) and circTFRC knockdown (sh-circTFRC) AGS cells following treatment with RSL3 (1 μM) in the absence or presence of ferrostatin-1 (0.75 μM) for 16 h. M Confocal microscopy showing the lipid ROS levels in control (sh-NC) and circTFRC knockdown (sh-circTFRC) AGS cells following treatment with RSL3 (1 μM) in the absence or presence of ferrostatin-1 (0.75 μM) for 16 h. Lipid ROS are stained with C11 BODIPY 581/591 probe (green). Nuclei are counterstained with DAPI (blue). Scale bar, 10 µm. Data are presented as mean ± SD. P-values were calculated using a two-tailed one-way ANOVA (A–F) or unpaired Student’s t test (H, J–L); **P < 0.01, ***P < 0.001, ****P < 0.0001. See also Fig. S4.

Article Snippet: Quantitative analysis of PI-positive populations was subsequently conducted using the Cytek® Northern LightsTM-CLC (NL-CLC) full spectrum flow cytometry system (Cytek® Biosciences, USA), with a minimum of 10,000 cells analyzed per condition.

Techniques: Knockdown, Staining, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Colony Assay, Transwell Assay, Migration, Confocal Microscopy, Two Tailed Test

Journal: Cell death & disease

Article Title: Circular RNA TFRC/SCD1 mRNA interaction regulates ferroptosis and metastasis in gastric cancer.

doi: 10.1038/s41419-025-07759-x

Figure Lengend Snippet: Fig. 6 CircTFRC exerted an oncogenic effect via SCD1 in GC cells. A CCK-8 assay showing the proliferation of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection at indicated time points (0, 24, 48, and 72 h). B, C Plate colony formation assay assessing colony formation in AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection over 10 days. D, E Transwell assay showing the migration of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection over 24 h. Scale bar, 100 µm. F PI staining showing the cell death rates of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection. G Flow cytometry showing the lipid ROS levels (stained with C11 BODIPY 581/591) of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection. ROS reactive oxygen species. H ELISA assays showing the relative MDA levels of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection. MAD malondialdehyde. The data are shown as the mean ± SD. The P values were determined by a two-tailed unpaired Student’s t test (C, E–H) or two-way ANOVA (A); **P < 0.01, ***P < 0.001, ****P < 0.0001. See also Fig. S7.

Article Snippet: Quantitative analysis of PI-positive populations was subsequently conducted using the Cytek® Northern LightsTM-CLC (NL-CLC) full spectrum flow cytometry system (Cytek® Biosciences, USA), with a minimum of 10,000 cells analyzed per condition.

Techniques: CCK-8 Assay, Control, Knockdown, Plasmid Preparation, Cotransfection, Colony Assay, Transwell Assay, Migration, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test