primary rabbit antibodies against p21  (Santa Cruz Biotechnology)

Journal: Cells

Article Title: EA.hy926 Cells and HUVECs Share Similar Senescence Phenotypes but Respond Differently to the Senolytic Drug ABT-263

doi: 10.3390/cells11131992

Figure Lengend Snippet: Doxorubicin induces the expression of senescence markers in a concentration-dependent manner in EA.hy926 cells and HUVECs. EA.hy926 human-endothelial-derived cells and HUVECs were treated with increasing concentrations of DOX (0.1 µM, 0.2 µM, and 0.5 µM) for 24 h. Thereafter, DOX was removed and cells were incubated in DOX-free media for 72 h. Then, cells were harvested and the total protein was extracted. Expression levels of senescence markers including p53, p21, and cyclin D1 in EA.hy926 cells (( A – C ), respectively) and HUVECs (( D – F ), respectively) were measured via Western blotting ( n = 4–6). Representative images of Western blots are shown. Values were normalized to α-tubulin and expressed relative to control cells. Values are presented as means ± SEM. Data were analyzed by one-way ANOVA followed by Dunnet’s multiple comparisons test. Note: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary mouse antibodies against p53 (catalog #2524) and primary rabbit antibodies against p21 (catalog #2947), cyclin D1 (catalog #2978), cleaved caspase-3 (catalog #9664), caspase-3 (catalog #9662), cleaved PARP (catalog #5625), PARP (catalog #9542), BCL-2 (catalog #4223), BCL-xL (catalog #2764), BCL-W (catalog #2724), BAK (catalog #12105), BAX (catalog #2774), and alpha-tubulin (catalog #2144) were purchased from Cell Signaling.

Techniques: Expressing, Concentration Assay, Derivative Assay, Incubation, Western Blot, Control

Journal: Molecular and Cellular Biochemistry

Article Title: In vitro study on effect of bardoxolone methyl on cisplatin-induced cellular senescence in human proximal tubular cells

doi: 10.1007/s11010-021-04295-y

Figure Lengend Snippet: Cisplatin treatment induces the increase in senescence-related gene transcriptions and cytokine secretion. The mRNA expression of senescence-related genes, MKI67 ( A ), CDKN2A ( B ), CDKN1A ( C ), CCND1 ( D ), IL-6 ( E ), and IL-8 ( F ), after cisplatin (20 µmol/L) treatment was determined by qPCR. Cytokine levels, IL-6 ( G ) and IL-8 ( H ), in cultured medium of cisplatin (20 or 50 µmol/L)-exposed cells were quantified by ELISA. n = 3, * P < 0.05, ** P < 0.01

Article Snippet: This was followed by incubation with horseradish peroxidase-conjugated secondary antibodies for 1 h. Primary antibodies against human p21 Waf1/Cip1 , p16 INK4a , phosphorylated H2AX (Ser139, γ-H2AX), retinoblastoma (Rb), phosphorylated Rb (Ser780, pRb), cyclin D, and caspase-3 were all purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA).

Techniques: Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Oncology reports

Article Title: MicroRNA-134-3p inhibits ovarian cancer progression by targeting flap structure-specific endonuclease 1 in vitro.

doi: 10.3892/or.2020.7844

Figure Lengend Snippet: Figure 2. miR-134-3p facilitates cell apoptosis and induces cell cycle arrest at the G0/G1 phase in SKOV-3 and OVCAR-3 cells. SKOV-3 and OVCAR-3 cells were transfected with miR-134-3p mimic or NC mimic. (A) TUNEL assay was performed to assess cell apoptosis (scale bar, 200 µm). (B) Western blot analysis was used to assess the protein expression levels of Blc-2, Bax, cleaved caspase-3 and cleaved caspase-9. (C) Flow cytometry assay was performed to assess the cell cycle. (D) Western blot analysis was used to assess the protein expression levels of cyclin D1, CDK2 and p21. The data are presented as the mean ± SD (n=3). *P<0.05 and **P<0.01 vs. NC mimic. PCNA, proliferating cell nuclear antigen; miR, microRNA; NC, negative control.

Article Snippet: The membranes were then blocked with TBS-Tween (TBST; 0.1% Tween-20) containing 1% skim milk powder at room temperature for 1 h, and then incubated with rabbit monoclonal primary antibodies against human p21 (1:1,000; cat. no. 2947S), cyclooxygenase-2 (Cox-2; 1:1,000; cat. no. 12282T), matrix metalloproteinase (MMP)2 (1:1,000; cat. no. 40994S), MMP9 (1:1,000; cat. no. 13667S), cyclin d1 (1:1,000; cat. no. 55506S), CdK2 (1:1,000; cat. no. 2546S), Bax (1:1,000; cat. no. 5023S), cleaved caspase-3 (1:1,000; cat. no. 9654S), cleaved caspase-9 (1:1,000; cat. no. 20750S), Bcl-2 (1:1,000; cat. no. 4223S), β-actin (1:1,000; cat. no. 4970T) and FEN1 (1:2,000; cat. no. 82354S) (all from Cell Signaling Technology, Inc.) at 4 ̊C overnight.

Techniques: Transfection, TUNEL Assay, Western Blot, Expressing, Flow Cytometry, Negative 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 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: 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