Journal: Journal of Extracellular Biology

Article Title: Extracellular Vesicles of Giardia duodenalis : Unravelling Their Virulence Factors and Potential to Induce Protection Against Experimental Giardiasis

doi: 10.1002/jex2.70155

Figure Lengend Snippet: Internalization of G. duodenalis TYI‐33‐EVs by intestinal epithelial cells (IEC‐6) . (A – C) TYI‐33–derived EVs were labeled with the membrane dye DiI for flow cytometry analysis and, in parallel experiments, co‐labeled with the membrane dye DiI and the RNA‐specific dye SYTO RNA. Labeled EVs were then incubated with IEC monolayers for 3 h at 37°C. A) Representative flow cytometry dot plots analyses illustrating EVs uptake by IECs. B) Quantification of Dil‐positive IECs, expressed as the percentage of cells exhibiting fluorescence, indicating EV internalization. C) Confocal fluorescence microscopy images showing IECs with internalized Dil‐labeled EVs (yellow) and associated RNA cargo stained with Syto RNA (green). Nuclei were counterstained with DAPI (blue). (D – F) Protein cargo of TYI‐33‐EVs was labeled with CFSE and incubated with IEC monolayers for 3 h at 37°C. D) Representative flow cytometry dot plots illustrating the uptake of CFSE‐labeled EVs by IECs. E) Quantification of CFSE‐positive IECs, expressed as the percentage of fluorescent cells, indicating internalization of EVs. F) Confocal fluorescence microscopy images showing IECs with internalized CFSE‐labeled EVs (green). Nuclei were counterstained with DAPI (blue). Experiments were performed in triplicate and statistical analysis was carried out using Mann–Whitney test.

Article Snippet: IEC‐6 cell line (ATCC Cat. No. CRL1592) was seeded on coverslips in the bottom of 24‐well plates (Sigma, Hamburg, Germany) and cultured in DMEM medium (GibcoBRL, Gaithersburg, MD, USA) supplemented with 10% FBS (HyClone, Marlborough, MA, USA), 0.1% glutamine and 1% antibiotic/antimycotic mixture (HyClone, Marlborough, MA, USA) at 37°C and 5% CO 2 .

Techniques: Derivative Assay, Labeling, Membrane, Flow Cytometry, Incubation, Fluorescence, Microscopy, Staining, MANN-WHITNEY

Journal: Journal of Extracellular Biology

Article Title: Extracellular Vesicles of Giardia duodenalis : Unravelling Their Virulence Factors and Potential to Induce Protection Against Experimental Giardiasis

doi: 10.1002/jex2.70155

Figure Lengend Snippet: Giardia duodenalis EVs induce cell death in intestinal epithelial cells (IEC‐6) . Scanning electron microscopy (SEM) images of IEC monolayers after 5 h of incubation at 37°C. A) Untreated control cells show an intact and confluent epithelial monolayer, with preserved morphology and junctional contacts. B) IECs exposed to live Giardia trophozoites display severe damage, including membrane blebbing and other surface irregularities. C) IECs incubated with 100 µg of total DMEM‐EVs protein exhibit morphological changes consistent with apoptotic processes, such as surface blebbing (arrow), membrane irregularities, and cell shrinkage. D) Cells treated with 100 µg of total TYI‐33‐EVs protein obtained in TYI‐33 culture exhibit pronounced structural alterations, such as intercellular separation (asterisk), blebbing, and cell shrinkage (arrow), morphological features consistent with apoptosis or stress‐induced cell death. The micrographs on the right shows a magnified view. Bar = 10 µm.

Article Snippet: IEC‐6 cell line (ATCC Cat. No. CRL1592) was seeded on coverslips in the bottom of 24‐well plates (Sigma, Hamburg, Germany) and cultured in DMEM medium (GibcoBRL, Gaithersburg, MD, USA) supplemented with 10% FBS (HyClone, Marlborough, MA, USA), 0.1% glutamine and 1% antibiotic/antimycotic mixture (HyClone, Marlborough, MA, USA) at 37°C and 5% CO 2 .

Techniques: Electron Microscopy, Incubation, Control, Membrane

Journal: Journal of Extracellular Biology

Article Title: Extracellular Vesicles of Giardia duodenalis : Unravelling Their Virulence Factors and Potential to Induce Protection Against Experimental Giardiasis

doi: 10.1002/jex2.70155

Figure Lengend Snippet: Localization of ZO‐1 and actin in intestinal epithelial cells (IEC‐6) after incubation with Giardia duodenalis TYI‐33‐EVs. A) IECs control, B) incubated with G. duodenalis live trophozoites or C) TYI‐33‐EVs. Preparations were analyzed with a confocal microscope considering xy ‐ and zy ‐planes. Arrows: ZO‐1 and polymerized actin are located at the cellular borders. Arrowheads: Incubation with either G. duodenalis trophozoites or TYI‐33‐EVs change their localization to the cytoplasm. Bar = 10 µm.

Article Snippet: IEC‐6 cell line (ATCC Cat. No. CRL1592) was seeded on coverslips in the bottom of 24‐well plates (Sigma, Hamburg, Germany) and cultured in DMEM medium (GibcoBRL, Gaithersburg, MD, USA) supplemented with 10% FBS (HyClone, Marlborough, MA, USA), 0.1% glutamine and 1% antibiotic/antimycotic mixture (HyClone, Marlborough, MA, USA) at 37°C and 5% CO 2 .

Techniques: Incubation, Control, Microscopy

Journal: Bioactive Materials

Article Title: Integrated fabrication of a shape-adaptable, antioxidative composite stent for effective closure and biological repair of enteroatmospheric fistula

doi: 10.1016/j.bioactmat.2026.01.014

Figure Lengend Snippet: Biocompatibility and cell migration of the composite stent. (A) Live/dead staining of IEC-6 cells cultured with the stent, showing cell viability. (B) Hemocompatibility of the stent: (i) hemolysis assay of red blood cells, (ii) corresponding hemolysis percentage. (C) Representative images of cell migration in the scratch assay. (D) Quantification of (i) migration area and (ii) percentage closure over time. (E) Representative Transwell images of migrated cells. (F) Quantification of migrated cells in the Transwell assay. Data were presented as mean ± SD (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Gel represents the G 2 S 4 hydrogel group, and Comp represents the composite stent group; the same abbreviations are used in the following figures.

Article Snippet: IEC-6 cells and RAW264.7 cells were purchased from KeyGEN BioTech (Nanjing, China).

Techniques: Migration, Staining, Cell Culture, Hemolysis Assay, Wound Healing Assay, Transwell Assay

Journal: Bioactive Materials

Article Title: Integrated fabrication of a shape-adaptable, antioxidative composite stent for effective closure and biological repair of enteroatmospheric fistula

doi: 10.1016/j.bioactmat.2026.01.014

Figure Lengend Snippet: Antioxidant and anti-inflammatory effects of the composite stent. (A) ROS levels in RAW264.7 and IEC-6 cells by DCFH-DA staining. (B, C) Quantification of ROS fluorescence by integrated density. (D) Mitochondrial ROS detected by MitoSOX staining. (E, F) Quantification of mitochondrial ROS by integrated density. (G) Co-culture system of the composite stent with macrophages using Transwell chambers. (H) Flow cytometry analysis of macrophage polarization based on CD86 (M1 marker) and CD206 (M2 marker) expression under different stimuli. (I) Quantitative analysis of CD86 + macrophages obtained from flow cytometry. (J–L) ELISA measurements of pro-inflammatory cytokines (J) TNF-α, (K) IL-6, and (L) IFN-β in the culture supernatant. Data were presented as mean ± SD (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: IEC-6 cells and RAW264.7 cells were purchased from KeyGEN BioTech (Nanjing, China).

Techniques: Staining, Fluorescence, Co-Culture Assay, Flow Cytometry, Marker, Expressing, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: CDK4/6 inhibitors enhance oxaliplatin efficacy in colorectal cancer with RB-dependent and tumor-selective activity in intestinal model

doi: 10.64898/2026.04.15.718743

Figure Lengend Snippet: (A) Representative fluorescence images of IEC-6 cells treated for 48 h with oxaliplatin (0.6 μM), abemaciclib (300 nM), palbociclib (400 nM), or their combinations. Viable cells were detected by Calcein AM fluorescence (green), and nuclei were counterstained with Hoechst (blue). Scale bar: 1000 μm. (B) Quantification of Calcein AM–positive cells following treatment with oxaliplatin and abemaciclib. (C) Quantification of Calcein AM–positive cells following treatment with oxaliplatin and palbociclib. (D) Quantification of ethidium homodimer–positive cells in IEC-6 cultures treated with oxaliplatin and abemaciclib. (E) Quantification of ethidium homodimer–positive cells in IEC-6 cultures treated with oxaliplatin and palbociclib. (F) A summary table showing the percentage of viable cells in HCT116 (tumor) and IEC-6 (non-tumor) cell lines across treatments. The data are presented as the mean ± SEM of three independent biological replicates. Quantification was performed by counting Calcein AM–positive cells relative to total nuclei. Statistical analysis was performed using one-way ANOVA followed by Šídák’s multiple comparisons test. Significance levels are indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: The non-tumoral rat intestinal epithelial cell line IEC-6 (ATCC: CRL-1592), kindly donated by Prof. José Garcia Abreu (ICB/UFRJ), was maintained in the same basal medium (DMEM/F-12 supplemented with 10% FBS) with additional insulin supplementation at a final concentration of 0.1 U/mL throughout all experimental procedures, including treatments.

Techniques: Fluorescence