Journal: Scientific Reports

Article Title: Neuroprotective effect of Ruminococcus albus on oxidatively stressed SH-SY5Y cells and animals

doi: 10.1038/s41598-017-15163-5

Figure Lengend Snippet: Effects of CRA-CM on the expression of SRF, C-fos and CDK2 in SH-SY5Y cells. The expression levels of SRF, C-fos and CDK2 were measured by western blotting and cropped blots are displayed ( A ), and the densities of SRF ( B ), c-Fos ( C ) and CDK2 ( D ) were quantified. The full-length blots are shown in Supplementary Fig. . The data are expressed as the mean ± SD of three independent experiments performed in triplicate.

Article Snippet: The membrane was incubated with 5% (w/v) skim milk in PBS (pH 7.4) containing 0.05% Tween-20 (PBS-T) at room temperature for 1 h, washed with PBS-T and then incubated with primary antibodies GAPDH (MA5-15738, endogenous control; 1:5,000 dilution, Thermo Fisher Scientific, Rockford, IL, USA), CDK2 (NB100-81842, 1:750 dilution, Novus Biologicals, Littleton, CO, USA) and Bax (633601, 1:500 dilution, Biolegend, San Diego, CA, USA) for 1 h at room temperature and against SRF (NBp1-33063, 1:500 dilution, Novus Biologicals) and c-Fos (AB1584, 1:500, Millipore) overnight at 4 °C with 5% (w/v) non-fat skim milk.

Techniques: Expressing, Western Blot

Journal: Cell Death Discovery

Article Title: ABCB1 confers resistance to carboplatin by accumulating stem-like cells in the G2/M phase of the cell cycle in p53 null ovarian cancer

doi: 10.1038/s41420-025-02435-7

Figure Lengend Snippet: A Cell cycle analysis in carR-SKOV3 and -OVCAR3 compared to each nonR group. (SKOV3: nonR; n = 16, carR; n = 13, OVCAR3: nonR; n = 10, carR; n = 12) Cell cycle distribution in each phase of the cell cycle was shown in graphs ( B ). Statistical analyses were conducted between nonR and carR cells that are distributed in the G2/M phase (SKOV3 nonR vs carR: p < 0.001/OVCAR3 nonR vs carR: p = 0.0042). C Immunoblotting analyses of G2/M transition related-proteins (phospho-cdc25c and CCNB1) in carR-SKOV3 and -OVCAR3 compared to each nonR group. Loading control: β-actin. Analysis of stemness profiling of each phase of the cell cycle in carR-SKOV3 (CD133: 1.55-fold, p < 0.001; LGR5: 1.79-fold, p = 0.005; CD117: 1.61-fold, p < 0.001; SOX2: 1.89-fold, p = 0.005) ( D ) and -OVCAR3 (CD133: p = 0.512; LGR5: p = 0.952; CD117: p = 0.839; SOX2: p = 0.082) ( E ) compared to nonR groups, respectively. F Flow cytometry plots showing the Ki67-positive cell proportion in carR-SKOV3 ( p = 0.0087) and -OVCAR3 ( p = 0.9469) compared to each nonR group. Comparison of Ki67-positive proportion in G2/M-gated fraction was displayed in graph ( G ). Data represent the means ± SD from multiple experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, NS not significant.

Article Snippet: After blocking procedure, membranes were incubated with primary antibodies against p53 (7F5), phospho-p53 (E9Y4U; Ser15), PARP1 (46D11), cleaved-PARP1 (D64E10; Asp214), BCL-XL (54H6), phospho-cdc25c (63F9; Ser216), cyclin B1 (D5C10), ΔNp73 (38C674.2; Novus, USA), and β-actin (all from Cell signaling, USA), respectively, and finally visualized by ECL solution (Thermo, USA).

Techniques: Cell Cycle Assay, Western Blot, Control, Flow Cytometry, Comparison

Journal: Cell Proliferation

Article Title: The mechanisms of nadroparin‐mediated inhibition of proliferation of two human lung cancer cell lines

doi: 10.1111/j.1365-2184.2012.00847.x

Figure Lengend Snippet: Western blot analysis of the effect of nadroparin on Cdk1 (a), p‐Cdk1 (b), Cdc25C (d) and p‐Cdc25C (e) protein levels and analysis of Cdk1 (c) and Cdc25C (f) mRNA expression in A549 cells. (a) Effect of nadroparin on Cdk1 protein levels. Blot is representative of three independent experiments. For densitometric analysis, resulting bands were quantified using IMAGEJ; data are mean ± SEM from all three experiments. **P < 0.01 for nadroparin (40 IU/ml) and colchicine‐treated versus ‐untreated cells (ANOVA); (b) Effect of nadroparin on p‐Cdk1 protein levels. The blot is representative of three independent experiments. For densitometric analysis, resulting bands were quantified using IMAGEJ; data are mean ± SEM from all three experiments. ***P < 0.0001 for nadroparin (40 IU/ml)‐treated versus ‐untreated cells (ANOVA); **P < 0.01 for colchicine‐treated versus ‐untreated cells (ANOVA); (c) Effect of nadroparin on Cdk1 mRNA expression. Data are mean ± SEM from three independent experiments. *P < 0.05 for nadroparin (40 IU/ml) and colchicine‐treated versus ‐untreated cells (ANOVA); (d) Effect of nadroparin on Cdc25C protein levels. Blot is representative of three independent experiments. For densitometric analysis, resulting bands were quantified using IMAGEJ; data are mean ± SEM from all three experiments. *P < 0.05 for nadroparin (40 IU/ml) and colchicine‐treated versus ‐untreated cells (ANOVA); (e) Effect of nadroparin on p‐Cdc25C protein levels. Blot is representative of three independent experiments. For densitometric analysis, resulting bands were quantified using IMAGEJ; data are mean ± SEM from all three experiments. **P < 0.01 for nadroparin (40 IU/ml) and colchicine‐treated versus ‐untreated cells (ANOVA); (f) Effect of nadroparin on Cdc25C mRNA expression. Data are mean ± SEM from three independent experiments. *P < 0.05 for nadroparin (40 IU/ml)‐treated versus ‐untreated cells (ANOVA); **P < 0.01 for colchicine‐treated versus ‐untreated cells (ANOVA).

Article Snippet: Mouse monoclonal antibodies against cyclin B1, Cdk1, Cdc25C and p21, goat anti‐mouse horseradish peroxidase labelled secondary antibody, anti‐human β‐actin and anti‐human α‐tubulin mouse monoclonal antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); rabbit monoclonal antibodies against p‐Cdk1 (Tyr15) and p‐Cdc25C (Ser216), anti‐human β‐actin rabbit monoclonal and goat anti‐rabbit horseradish peroxidase labelled secondary antibody were purchased from Histo‐Line Laboratories (Epitomics, Milan, Italy).

Techniques: Western Blot, Expressing