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

Journal: Frontiers in Pharmacology

Article Title: Hydroxysafflor yellow A attenuates sepsis-induced intestinal barrier dysfunction by modulating Bcl-2/SOD2-mediated mitochondrial apoptosis

doi: 10.3389/fphar.2026.1728183

Figure Lengend Snippet: Proteomic Analysis for Identifying Key Molecules Mediating Ferroptosis in Sepsis-Associated IEC-6 Cells (A) KEGG pathway enrichment analysis. (B) Immunofluorescence staining of ZO-1 (scale bar = 50 μm) and Western blot analysis of ZO-1 expression. (C) CCK-8 assay of cell viability (n = 8). (D) DCFH-DA showing ROS levels (scale bar = 20 μm, n = 8) (E) TUNEL staining of apoptotic cells. (F) Ki-67 immunofluorescence of proliferating cells (G) MitoTracker Red CMXRos staining showing mitochondrial morphology (scale bar = 50 μm, n = 3). (H) TEM images of mitochondrial ultrastructure. (I) Impact of LPS on mitochondrial respiration in IEC-6 cells (n = 3).

Article Snippet: The rat intestinal epithelial cell line IEC-6 (ATCC, USA) was maintained in Dulbecco’s Modified Eagle Medium (DMEM; Hyclone, USA) supplemented with 10% fetal bovine serum (FBS; Hyclone, USA) under standard conditions (37 °C, 5% CO 2 , humidified incubator).

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, CCK-8 Assay, TUNEL Assay

Journal: Frontiers in Pharmacology

Article Title: Hydroxysafflor yellow A attenuates sepsis-induced intestinal barrier dysfunction by modulating Bcl-2/SOD2-mediated mitochondrial apoptosis

doi: 10.3389/fphar.2026.1728183

Figure Lengend Snippet: Molecular Docking, Molecular Dynamics Analysis, and in vitro / in vivo Validation of Key Proteins. (A) Chemical structure of HSYA. (B) Venn diagram integrating sepsis-, HSYA-, and apoptosis-related targets, highlighting Bcl-2 and SOD2 as key molecules. (C) Molecular docking and molecular dynamics analysis showing binding interactions of HSYA with Bcl-2. (D) Molecular docking and molecular dynamics analysis showing binding interactions of HSYA with SOD2. (E) Western blot analysis of Bcl-2 and SOD2 protein expression in IEC-6 cells (n = 3). (F) Western blot analysis of Bcl-2 and SOD2 protein expression in intestinal tissues (n = 3).

Article Snippet: The rat intestinal epithelial cell line IEC-6 (ATCC, USA) was maintained in Dulbecco’s Modified Eagle Medium (DMEM; Hyclone, USA) supplemented with 10% fetal bovine serum (FBS; Hyclone, USA) under standard conditions (37 °C, 5% CO 2 , humidified incubator).

Techniques: In Vitro, In Vivo, Biomarker Discovery, Binding Assay, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Hydroxysafflor yellow A attenuates sepsis-induced intestinal barrier dysfunction by modulating Bcl-2/SOD2-mediated mitochondrial apoptosis

doi: 10.3389/fphar.2026.1728183

Figure Lengend Snippet: In vitro effects of HSYA on IEC-6 cells following Bcl-2 inhibition. (A) Western blot analysis showing the relative expression level of Bcl-2 after treatment in Si-Bcl-2 of IEC-6 cells (n = 3). (B) Western blot analysis showing the effects of HSYA + Si-Bcl2 on the expression of Bcl2 in LPS-stimulated IEC-6 cells (n = 3). (C) TUNEL staining of apoptotic cells (scale bar = 50 μm, n = 3). (D) Ki-67 immunofluorescence for proliferating cells. (E,F) DCFH-DA (n = 3) and MitoTracker Red CMXRos staining (n = 8) showing intracellular ROS levels and mitochondrial morphology in IEC-6 cells (scale bar = 20 μm). (G) TEM images showing mitochondrial ultrastructure in IEC-6 cells following HSYA and HSYA + Si-Bcl-2 treatment. (H) Impact of HSYA + Si-Bcl-2 treatment on mitochondrial respiration in LPS-stimulated IEC-6 cells (n = 3).

Article Snippet: The rat intestinal epithelial cell line IEC-6 (ATCC, USA) was maintained in Dulbecco’s Modified Eagle Medium (DMEM; Hyclone, USA) supplemented with 10% fetal bovine serum (FBS; Hyclone, USA) under standard conditions (37 °C, 5% CO 2 , humidified incubator).

Techniques: In Vitro, Inhibition, Western Blot, Expressing, TUNEL Assay, Staining, Immunofluorescence

Journal: Frontiers in Pharmacology

Article Title: Hydroxysafflor yellow A attenuates sepsis-induced intestinal barrier dysfunction by modulating Bcl-2/SOD2-mediated mitochondrial apoptosis

doi: 10.3389/fphar.2026.1728183

Figure Lengend Snippet: Effects of HSYA on intestinal epithelial cells under silence SOD-2. (A,B) Western blot analysis of SOD2 protein expression in IEC-6 cells (n = 3). (C) TUNEL staining for apoptotic cells (scale bar = 50 μm). (D) Ki-67 immunofluorescence for proliferating cells (scale bar = 50 μm, n = 8). (E,F) DCFH-DA (n = 8)and MitoTracker Red CMXRos staining (n = 3) showing intracellular ROS accumulation and mitochondrial morphology in IEC-6 cells (scale bar = 20 μm). (G) TEM images showing mitochondrial ultrastructure in IEC-6 cells treated with HSYA or HSYA + Si-SOD2 (scale bar = 1 μm). (H) Impact of HSYA + Si-SOD2 treatment on mitochondrial respiration in LPS-stimulated IEC-6 cells (n = 3).

Article Snippet: The rat intestinal epithelial cell line IEC-6 (ATCC, USA) was maintained in Dulbecco’s Modified Eagle Medium (DMEM; Hyclone, USA) supplemented with 10% fetal bovine serum (FBS; Hyclone, USA) under standard conditions (37 °C, 5% CO 2 , humidified incubator).

Techniques: Western Blot, Expressing, TUNEL Assay, Staining, Immunofluorescence

Journal: Scientific Reports

Article Title: Toxic effects of Helix aspersa snail egg hydrolyzates obtained by static in vitro digestion on Caco-2 colorectal adenocarcinoma cells

doi: 10.1038/s41598-025-11605-7

Figure Lengend Snippet: The integrity of the plasma membrane of Caco-2 cells incubated with hydrolyzates from Helix aspersa maxima and Helix aspersa aspersa eggs (concentrated and diluted 10, 100, and 1000 times) for ( a ) 24 h and ( b ) 72 h; and integrity of plasma membrane of IEC-6 cells treated with these hydrolyzates for ( c ) 24 h and ( d ) 72 h. H, digestive fluids; MN and AN, non-digested extracts from H. a. maxima and H. a. aspersa eggs, respectively; MH and AH, hydrolyzates from H. a. maxima and H. a. aspersa eggs, respectively; LDH, lactate dehydrogenase. Error bars show the standard error of the mean. An asterisk (*) indicates values that differ from control (for cells treated with H of appropriate concentration) at p < 0.05, two asterisks (**) - at p < 0.01, and three asterisks (***) – at p < 0.001. n = 6.

Article Snippet: A human epithelial colorectal adenocarcinoma cell line (Caco-2; the European Collection of Authenticated Cell Cultures, Sigma-Aldrich, St. Louis, MO, USA; 55 passage) and a control rat intestinal epithelial cell line (IEC-6; the American Type Culture Collection, Manassas, VA, USA; 17 passage) were cultured in 96-well polystyrene plates intended for adherent culture, with an initial concentration of 1 × 10 4 cells/100 μL of medium, for the test described in “Effect of hydrolyzates on the integrity of plasma membranes of Caco-2 and IEC-6 cells” section.

Techniques: Clinical Proteomics, Membrane, Incubation, Control, Concentration Assay