Journal: Oncology Reports

Article Title: Suppression of CDCA3 inhibits prostate cancer progression via NF-κB/cyclin D1 signaling inactivation and p21 accumulation

doi: 10.3892/or.2021.8253

Figure Lengend Snippet: Knockdown of CDCA3 induces G0/G1-phase arrest in DU145 and PC-3 cells. (A) Flow cytometry was used to analyze the cell cycle following CDCA3 knockdown in DU145 and PC-3 cells. (B) Cell cycle-associated protein levels, including cyclin D1 and p21, were analyzed via western blotting. GAPDH was used as a loading control. Data are presented as the mean ± SD of three independent experiments. **P<0.01 and ***P<0.001. CDCA3, cell division cycle-associated 3; sh-, short hairpin; NC, negative control.

Article Snippet: The following antibodies were used: Mouse monoclonal anti-GAPDH (1:2,000; cat. no. 33033M; BIOSS), rabbit polyclonal anti-CDCA3 (1:1,000; cat. no. YT0819), rabbit polyclonal anti-cleaved caspase-3 (1:1,000; cat. no. YC0006), rabbit polyclonal anti-pro-caspase-3 (1:1,000; cat. no. YT6113), and rabbit polyclonal anti-cyclin-dependent kinase inhibitor 1 (p21; 1:1,000; cat. no. YT3497; all from Immunoway Biotechnology Company), rabbit polyclonal anti-cyclin D1 (1:1,000; cat. no. 0623R; BIOSS), mouse monoclonal anti-NFκB-p65 (1:1,000; cat. no. YM311), rabbit polyclonal anti-phosphorylated (p)-NFκB-p65 (1:1,000; cat. no. YP0192), rabbit polyclonal anti-IKKα/β (1:1,000; cat. no. YT2302), and rabbit polyclonal anti-NFκB-p105/p50 (1:1,000; cat. no. YT3101; all from Immunoway Biotechnology Company), HRP-labeled goat anti-rabbit secondary antibody (1:5,000; cat. no. 40295G-HRP; BIOSS) and HRP-labeled goat anti-mouse secondary antibody (1:5,000; cat. no. 0368G-HRP; BIOSS).

Techniques: Flow Cytometry, Western Blot, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Aspirin promotes apoptosis and inhibits proliferation by blocking G 0 /G 1 into S phase in rheumatoid arthritis fibroblast-like synoviocytes via downregulation of JAK/STAT3 and NF-κB signaling pathway

doi: 10.3892/ijmm.2018.3883

Figure Lengend Snippet: Effects of aspirin on Bcl-2, BAX, PARP1, Cyclin D1 and P21 expression in RA-FLS. (A) After RA-FLS were exposed to various concentration of aspirin for 24 h, western blotting was used to determine Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein levels. The corresponding internal control was GADPH. Changes in Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein expression in RA-FLS treated with aspirin are shown: Bcl-2, PARP1, Cyclin D1 and P21 were decreased while BAX was increased, each with increasing aspirin concentration. (B) Bar graph shows the gray value analysis of Bcl-2, BAX, PARP1, Cyclin D1 and P21. Data are presented as the means ± SD (error bars) from three independent experiments. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 vs. DMSO group. We set the value as target protein vs. GAPDH, and set the value of control group as 1. CCND1, Cyclin D1; RA-FLS, rheumatoid arthritis-fibroblast-like synoviocytes; Bcl-2, B-cell lymphoma-2; BAX, Bcl-2-associated X protein; PARP1, poly (ADP-ribose) polymerase 1.

Article Snippet: The membranes were blocked with 5% BSA in TBS-T for 1 h, and then incubated with rabbit anti-Bax (1:2,000 dilution, Proteintech Group; #S0599-2-lg), anti-Bcl-2 (1:1,000 dilution, Santa Cruz Biotechnology, Inc.; #sc-7382), anti-PARP1 (1:2,000 dilution, Proteintech Group; #13371-1-AP), anti-P21 (1:1,000 dilution, #EPR3993), anti-cyclin D1 (1:10,000 dilution, #EPR2241; both from Abcam), anti-P65 (1:2,000 dilution; #8242), anti-p-P65 (1:2,000 dilution; #3033; both from Cell Signaling Technology, Inc.), anti-P50/P105 (1:1,000 dilution, Abcam: #E381), anti-p-P50/105 (1:1,000 dilution, Affinity Company: #AF3219), anti-STAT3 (1:2,000 dilution, #EPR787Y) and anti-p-STAT3 (1:200,000 dilution, #EP2147Y; both from Abcam) antibodies for 2 h at room temperature.

Techniques: Expressing, Concentration Assay, Western Blot

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