cck8 working solution  (Dojindo Labs)

Journal: Lipids in health and disease

Article Title: Fucoidan ameliorates lipid accumulation, oxidative stress, and NF-κB-mediated inflammation by regulating the PI3K/AKT/Nrf2 signaling pathway in a free fatty acid-induced NAFLD spheroid model.

doi: 10.1186/s12944-025-02483-z

Figure Lengend Snippet: Fig. 1 Fucoidan reduced FFA-induced lipid accumulation in HepG2 cells. (A) The experimental flow of FFAs stimulation and fucoidan treatment in HepG2 cells. HepG2 cells were adhered in plates and cultured for 24 h, then stimulated with FFA (0.5 mM) for 24 h, further treated with DMSO (control) or fucoi dan (1, 10, 50, 100 µg/ml) for additional 24 h. (B) Detection of fucoidan cytotoxicity in HepG2 cells. HepG2 cells were stimulated with DMSO or fucoidan (1, 10, 50, 100 µg/ml) for 24 h, then the cell viability was detected using the CCK-8 assay. (C) Fucoidan improved FFA-induced cytotoxicity. HepG2 cells were treated as described in (A), and then the cell viability was detected using the CCK-8 assay. (D) Fucoidan decreased intracellular TG content in HepG2 cells. HepG2 cells were treated as described in (A), and then the intracellular TG content was measured. (E) Fucoidan inhibited lipid droplet formation in HepG2 cells. HepG2 cells were treated as described in (A), and then the cells were stained with oil red O. Data were analyzed by the student’s t test and expressed as the mean ± SD (n = 3). * represents the differences compared to the control group. # represents the differences compared to the DMSO group. ** P < 0.01; ##P < 0.01

Article Snippet: HepG2 cells were cultured in a 96-well plate with complete medium and stimulated with DMSO or fucoidan (1, 10, 50, 100 μg/mL) for 24 h. After that, CCK8 working solution (MedChemExpress, USA) was added into each well and incubated at 37 °C for 30 min.

Techniques: Cell Culture, Control, CCK-8 Assay, Staining

Journal: Antioxidants

Article Title: Large Fibrous Connective Tissue Reduces Oxidative Stress to Form a Living Cell Scaffold in Adipose Grafts

doi: 10.3390/antiox14030270

Figure Lengend Snippet: Characterization of adipocytes and adipose-derived stem cells (ADSCs) in areas adjacent (ALFC) and distant (DLFC) to large fibrous connective tissue (LFC) (n = 5, unpaired t -test). ( A , B ) Perilipin immunofluorescence of adipose tissue after 3 days and 7 days of incubation in serum-free medium under 1% volume fraction oxygen. ( C , D ) Dedifferentiation of mature adipocytes after 14 days and 21 days of ceiling culture. ( E ) Proliferative capacity of ADSCs by CCK-8 test. ( F , G ) Cell migration ability of ADSCs within 24 h. ( H ) Chondrogenic, osteogenic, and adipogenic differentiation potential of ADSCs under normoxic (21% O 2 ) and hypoxic (1% O 2 ) conditions. ( I ) Comparison of adipogenic area stained with Oil Red O. *, p < 0.05. ns: Not significant. Scale bar: 100 μm ( A ); 10 μm (( C ) (200×)); 20 μm (( C ) (100×)); 20 μm ( F ); 20 μm ( H ).

Article Snippet: CCK8 working solution (100 μL, CCK8:DMEM = 1:9) was added and incubated at 37 °C for 2 h. The OD value at a wavelength of 450 nm was measured by ELISA (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Derivative Assay, Immunofluorescence, Incubation, CCK-8 Assay, Migration, Comparison, Staining

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Deoxycholic acid aggravates necrotizing enterocolitis through downregulation of mesenchymal-epithelial transition factor expression

doi: 10.1590/1414-431X2024e14046

Figure Lengend Snippet: Elevated deoxycholic acid (DCA) levels in a mouse model of necrotizing. enterocolitis (NEC). A and B , Survival rate and weight of the mice on the final day of the modeling procedure ( A , log-rank; B , unpaired t -tests). C , Representative photomicrographs of ileum tissue from each group (n=6; magnification ×10 and 20×, with the scale bars indicating 100 and 50 µm, respectively). D , Histopathological scoring of ileum tissue in mice (on a 0-4 scale; n=6; Kruskal-Wallis test). E and F , Quantification of bile acids in serum and fecal samples (n≥4; unpaired t -tests). G , IEC-6 cells treated with DCA for 24 h, and cell proliferation assessed using the CCK8 assay (n=3; unpaired t -tests). H and I , Migration rate of IEC-6 cells assessed by wound-healing assay after treatment with 200 µM DCA for 24 h (n=6; unpaired t -tests; scale bar indicates 100 µm). The data are reported as means±SD. *P<0.05, **P<0.01, ***P<0.001,****P<0.0001.

Article Snippet: Following a 24-h incubation period, 10 μL of CCK8 working solution (Dojindo, Japan) was added to each well, followed by further incubation at 37°C for 4 h. The absorbance was then measured at 450 nm using an enzyme-linked immunosorbent assay reader (PerkinElmer, USA).

Techniques: CCK-8 Assay, Migration, Wound Healing Assay

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Deoxycholic acid aggravates necrotizing enterocolitis through downregulation of mesenchymal-epithelial transition factor expression

doi: 10.1590/1414-431X2024e14046

Figure Lengend Snippet: Deoxycholic acid (DCA) suppressed the proliferation and migration of intestinal epithelial cells via mesenchymal-epithelial transition (MET) factor inhibition. A and B , IEC-6 cells were treated with 200 µM DCA for 24 h, followed by western blot analysis to detect MET expression. C-F , The expression levels of MET, p-STAT3, and STAT3 were measured in MET-deficient IEC-6 cells by western blot analysis. G , Proliferation rate of control and MET-deficient IEC-6 cells assessed by a CCK8 assay. H and I , The migration capacity of MET-deficient IEC-6 cells was evaluated using a wound-healing assay. Scale bar indicates 100 µm. The data are reported as means±SD (n=3). **P<0.01 and ****P<0.0001, unpaired t -tests. ns: non-significant.

Article Snippet: Following a 24-h incubation period, 10 μL of CCK8 working solution (Dojindo, Japan) was added to each well, followed by further incubation at 37°C for 4 h. The absorbance was then measured at 450 nm using an enzyme-linked immunosorbent assay reader (PerkinElmer, USA).

Techniques: Migration, Inhibition, Western Blot, Expressing, Control, CCK-8 Assay, Wound Healing Assay