Journal: Antioxidants (Basel, Switzerland)

Article Title: Inhibition of Oxidative Stress and ALOX12 and NF-κB Pathways Contribute to the Protective Effect of Baicalein on Carbon Tetrachloride-Induced Acute Liver Injury.

doi: 10.3390/antiox10060976

Figure Lengend Snippet: Figure 2. Protein expression levels involved ferroptosis were detected in the liver tissues of mice challenged with CCl4. (A) Protein expression levels of ALOX12, HO-1, COX-2, p21 and Nrf2 at 6 and 24 h identified by Western blotting analysis of liver tissues of mice treated with CCl4; the quantitative analysis is showed in (B) (n = 4). 6 h vs. ctrl, * p < 0.05, ** p < 0.01; 24 h vs. ctrl, # p < 0.05, ## p < 0.01. Ctrl: control.

Article Snippet: The following primary antibodies were used to probe the membranes: primary rabbit antibodies against p21 (1:1000), COX2 (1:1000), Nrf2 (1:1000), and HO-1 (1:1000) (ProteinTech Group, Inc., Chicago, IL, USA), mouse monoclonal antibody against ALOX12 (1:1000) (Abcam, Cabridge, MA, USA) and β-actin (1:1000) (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Western Blot, Control

Journal: Antioxidants (Basel, Switzerland)

Article Title: Inhibition of Oxidative Stress and ALOX12 and NF-κB Pathways Contribute to the Protective Effect of Baicalein on Carbon Tetrachloride-Induced Acute Liver Injury.

doi: 10.3390/antiox10060976

Figure Lengend Snippet: Figure 6. Effects of baicalein supplementation on apoptotic pathway in the liver tissues of mice challenged with CCl4. (A) Representative TUNEL-stained sections showing apoptosis in the liver tissue of mice. (B) Quantitative analysis of TUNEL positive rates (n = 4). (C,D) Activities of (C) caspases-3 and -9 (D) (n = 6). (E) The relative expression of Bax, GADD45a and p21 mRNAs in the liver tissues (n = 5). ** p < 0.01, compared to the control group; # p < 0.05 or ## p < 0.01, compared to the CCl4 only group. Bar = 100 µm. Bai: baicalein.

Article Snippet: The following primary antibodies were used to probe the membranes: primary rabbit antibodies against p21 (1:1000), COX2 (1:1000), Nrf2 (1:1000), and HO-1 (1:1000) (ProteinTech Group, Inc., Chicago, IL, USA), mouse monoclonal antibody against ALOX12 (1:1000) (Abcam, Cabridge, MA, USA) and β-actin (1:1000) (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: TUNEL Assay, Staining, Expressing, Control

Journal: Journal of toxicologic pathology

Article Title: AMPK-mTOR-ULK1-mediated autophagy protects carbon tetrachloride-induced acute hepatic failure by inhibiting p21 in rats.

doi: 10.1293/tox.2020-0022

Figure Lengend Snippet: Fig. 3. Autophagy inhibition enhances the level of p21. Cyclin-dependent kinase inhibitor p21 was observed by immunofluorescence after carbon tetrachloride (CCl4) treatment in the presence or absence of chloroquine (CQ) (A). The number of ~300 cells with punctate FITC- p21 is displayed as a histogram (B). All data are represented as the mean ± SD (n=4) and analyzed by one-way ANOVA with SPSS 19.0. **P<0.01 compared with the control group. #P<0.05, ##P<0.01, compared to the respective CCl4 group. The oblique lines represent the p21 puncta. Scale bar: 100 μm.

Article Snippet: Sections were fixed with 4% paraformaldehyde (Sinopharm Chemical Reagent Co., Ltd, Shanghai, China) at 4°C for 30 min and then permeabilized with 1% Triton X-100 for 20 min. After three washes with PBS, the slices were blocked with 5% BSA blocking solution and incubated with LC3B (cat. no. 3868) and p21 (cat. no. 2947) antibodies overnight at 4°C; these antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Inhibition, Immunofluorescence, Control

Journal: Journal of toxicologic pathology

Article Title: AMPK-mTOR-ULK1-mediated autophagy protects carbon tetrachloride-induced acute hepatic failure by inhibiting p21 in rats.

doi: 10.1293/tox.2020-0022

Figure Lengend Snippet: Fig. 5. Schematic of the association between autophagy and carbon tetrachloride (CCl4)-induced acute hepatic failure (AHF). CCl4-induced AHF causes exhaustion of hepatic energy, which subsequently activates AMPKα, facilitates the inacti- vation of mTORC1, promotes the dissociation of the ULK1 compound and initiates the occurrence of autophagy. Au- tophagy can serve an important protective function against hepatic injury by inhibiting p21.

Article Snippet: Sections were fixed with 4% paraformaldehyde (Sinopharm Chemical Reagent Co., Ltd, Shanghai, China) at 4°C for 30 min and then permeabilized with 1% Triton X-100 for 20 min. After three washes with PBS, the slices were blocked with 5% BSA blocking solution and incubated with LC3B (cat. no. 3868) and p21 (cat. no. 2947) antibodies overnight at 4°C; these antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: PloS one

Article Title: Sterigmatocystin-induced DNA damage triggers G2 arrest via an ATM/p53-related pathway in human gastric epithelium GES-1 cells in vitro.

doi: 10.1371/journal.pone.0065044

Figure Lengend Snippet: Figure 4. The p53-p21 pathway is activated in ST-treated GES-1 cells. GES-1 cells were treated with different concentrations of ST (0.075, 0.3, 1.5, and 3 mM) or solvent for 48 h. (A) Representative immunoblots show the effect of ST treatment on the phosphorylation of p53 (Ser-15) and the expression of p53 and p21. b-actin was used as the normalization control. (B) Intensities of the immunoreactive bands were quantified by densitometric scanning and compared with those of the control (considered ‘‘1’’). The values shown represent the means 6 SD. *P,0.05 compared with the solvent-treated control group. doi:10.1371/journal.pone.0065044.g004

Article Snippet: The primary antibodies used for the Western blot analysis were mouse anti-human Cyclin B1 antibody (eBioscience, CA, USA), rabbit anti-human Cdc2, Cdc25C, ATM, phospho-ATM (Ser-1981), and phospho-Chk2 (Thr-68) monoclonal antibodies (Epitomics, CA, USA), rabbit anti-human Chk2 monoclonal antibodies (Millipore, MA, USA), rabbit anti-human phospho-Cdc2 (Tyr15) and phospho-Cdc25C (Ser216) monoclonal antibodies (Cell Signaling Technology, MA, USA), mouse antihuman phospho-p53 (Ser15) monoclonal antibody (Cell Signaling Technology, MA, USA), rabbit anti-human p53, Bax, and caspase-3 antibodies, and mouse anti-human p21 waf1 and Bcl-2 antibodies (Santa Cruz, CA, USA).

Techniques: Solvent, Western Blot, Phospho-proteomics, Expressing, Control

Journal: PloS one

Article Title: Sterigmatocystin-induced DNA damage triggers G2 arrest via an ATM/p53-related pathway in human gastric epithelium GES-1 cells in vitro.

doi: 10.1371/journal.pone.0065044

Figure Lengend Snippet: Figure 6. Silencing of p53 by specific p53 siRNA inhibited ST-induced G2 arrest. Cells were either not transfected or transfected with 100 nM p53 siRNA and then treated with 3 mM ST for 48 h. (A) Cells were subjected to immunoblot analysis for p-p53 (Ser15), p53, p21, and (C) the regulators related to G2 arrest. NC: cells transfected with the same concentration of negative control siRNA. b-actin was used as the loading control. (B, D) Intensities of the immunoreactive bands in ‘‘A’’ and ‘‘C’’ were quantified by densitometric scanning and compared with those of the control (considered ‘‘1’’). (E) The cell cycle phases of the cells were analyzed by FCM. The values shown represent the means 6 SD, *P,0.05 compared with the solvent-treated control group. mP,0.05 compared with the ST-treated groups. #P,0.05 compared with the p53 siRNA-treated groups. doi:10.1371/journal.pone.0065044.g006

Article Snippet: The primary antibodies used for the Western blot analysis were mouse anti-human Cyclin B1 antibody (eBioscience, CA, USA), rabbit anti-human Cdc2, Cdc25C, ATM, phospho-ATM (Ser-1981), and phospho-Chk2 (Thr-68) monoclonal antibodies (Epitomics, CA, USA), rabbit anti-human Chk2 monoclonal antibodies (Millipore, MA, USA), rabbit anti-human phospho-Cdc2 (Tyr15) and phospho-Cdc25C (Ser216) monoclonal antibodies (Cell Signaling Technology, MA, USA), mouse antihuman phospho-p53 (Ser15) monoclonal antibody (Cell Signaling Technology, MA, USA), rabbit anti-human p53, Bax, and caspase-3 antibodies, and mouse anti-human p21 waf1 and Bcl-2 antibodies (Santa Cruz, CA, USA).

Techniques: Transfection, Western Blot, Concentration Assay, Negative Control, Control, Solvent

Journal: Integrative cancer therapies

Article Title: Scutellaria Barbata D Don Inhibits Colorectal Cancer Growth via Suppression of Multiple Signaling Pathways.

doi: 10.1177/1534735413508811

Figure Lengend Snippet: Figure 3. Effect of ethanol extract of Scutellaria barbata D Don (EESB) on cell proliferation and the expression of Cyclin D1, CDK4, and p21 in colorectal cancer (CRC) xenograft mice. (A) At the end of the study, tumor tissues were processed for immunohistochemical (IHC) staining for proliferating cell nuclear antigen (PCNA). The photographs are representative images taken at a magnification of 400×. Quantification of IHC assay was represented as percentage of positively stained cells. Data shown are mean ± standard deviation (error bars) of 6 individual mice in each group. *P < .05, versus controls. (B, C) The mRNA or protein expression levels of Cyclin D1, CDK4, and p21 were determined by reverse transcriptase–polymerase chain reaction (RT-PCR) and Western blot analyses. GAPDH and β-actin were used as the internal controls for the RT-PCR and Western blotting, respectively. Images are representatives of 3 individual mice in each group. (D, E) Quantitation of mRNA and protein expression was assayed by densitometric analysis. The data were normalized to the mean mRNA or protein expression of untreated control (100%). *P < .01, versus controls.

Article Snippet: Phospho-STAT3 (p-STAT3), total STAT3, Bcl-2, Bax, p21, cyclinD1, CDK4 antibodies, horseradish peroxidase (HRP)–conjugated secondary antibodies were obtained from Cell Signaling (Beverly, MA).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry, Staining, Standard Deviation, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Quantitation Assay, Control