Journal: Aging Cell

Article Title: Advanced maternal age causes premature placental senescence and malformation via dysregulated α‐Klotho expression in trophoblasts

doi: 10.1111/acel.13417

Figure Lengend Snippet: α‐Klotho regulates the genes of the CAM pathway in JAR cells. (a) Volcano plot of the significant differences in gene expression levels between Sh‐KL and Sh‐NC JAR cells; genes showing the greatest differences were analyzed by (b) KEGG and (c) GO analysis; (d‐e) mRNA levels and protein levels of CDH4, CLDN3, and ITGAM were examined separately by RT‐qPCR and Western blotting in various JAR cells, n = 3 per group; (f‐g) mouse placentas collected at different gestational ages: GD14.5 (Ad‐Ctrl, n = 6; Ad‐Klotho, n = 6) and GD18.5 (Ad‐Ctrl, n = 5; Ad‐Klotho, n = 5); (h‐i) placentas from young and aged mice on GD18.5, n = 6; (j‐k) human term placentas, protein levels ( n = 6 per group), gene level, n = 7/8; and (l‐m) human first trimester villi, protein levels ( n = 6 per group), gene level, n = 7/8. All data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. Mann–Whitney U test or one‐way ANOVA. NS, nonsignificant. All experiments were performed in triplicate.

Article Snippet: Next, the sections were incubated with rabbit mAb against α‐Klotho (1:100; AG27759; Proteintech, Rosemont, IL, USA), rabbit mAb against CLDN3 (1:100; AG9411; Proteintech), rabbit mAb against CDH4 (1:400; Proteintech), rabbit mAb against ITGAM (1:200; AG16327; Proteintech), mouse mAb against cytokeratin 7 (CK7) (1:100; EPR1619Y; Abcam, Cambridge, UK), mouse mAb against human leukocyte antigen G (HLA‐G) (1:100; MEM‐G/1; Proteintech), and rabbit mAb against CD31 (1:400; D8V9E; Cell Signaling Technology, USA) at 4°C overnight, followed by treatment with a secondary antibody conjugated with horseradish peroxidase for 30 min at RT.

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, MANN-WHITNEY

Journal: Aging Cell

Article Title: Advanced maternal age causes premature placental senescence and malformation via dysregulated α‐Klotho expression in trophoblasts

doi: 10.1111/acel.13417

Figure Lengend Snippet: α‐Klotho regulates the genes of the CAM pathway in JAR cells. (a) Volcano plot of the significant differences in gene expression levels between Sh‐KL and Sh‐NC JAR cells; genes showing the greatest differences were analyzed by (b) KEGG and (c) GO analysis; (d‐e) mRNA levels and protein levels of CDH4, CLDN3, and ITGAM were examined separately by RT‐qPCR and Western blotting in various JAR cells, n = 3 per group; (f‐g) mouse placentas collected at different gestational ages: GD14.5 (Ad‐Ctrl, n = 6; Ad‐Klotho, n = 6) and GD18.5 (Ad‐Ctrl, n = 5; Ad‐Klotho, n = 5); (h‐i) placentas from young and aged mice on GD18.5, n = 6; (j‐k) human term placentas, protein levels ( n = 6 per group), gene level, n = 7/8; and (l‐m) human first trimester villi, protein levels ( n = 6 per group), gene level, n = 7/8. All data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. Mann–Whitney U test or one‐way ANOVA. NS, nonsignificant. All experiments were performed in triplicate.

Article Snippet: Next, the sections were incubated with rabbit mAb against α‐Klotho (1:100; AG27759; Proteintech, Rosemont, IL, USA), rabbit mAb against CLDN3 (1:100; AG9411; Proteintech), rabbit mAb against CDH4 (1:400; Proteintech), rabbit mAb against ITGAM (1:200; AG16327; Proteintech), mouse mAb against cytokeratin 7 (CK7) (1:100; EPR1619Y; Abcam, Cambridge, UK), mouse mAb against human leukocyte antigen G (HLA‐G) (1:100; MEM‐G/1; Proteintech), and rabbit mAb against CD31 (1:400; D8V9E; Cell Signaling Technology, USA) at 4°C overnight, followed by treatment with a secondary antibody conjugated with horseradish peroxidase for 30 min at RT.

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, MANN-WHITNEY

Journal: Poultry Science

Article Title: Marine algal polysaccharides alleviate aflatoxin B1-induced bursa of Fabricius injury by regulating redox and apoptotic signaling pathway in broilers

doi: 10.1016/j.psj.2020.10.050

Figure Lengend Snippet: Primers used for quantitative real-time PCR.

Article Snippet: Then, it was transferred to nitrocellulose membranes using a tank transfer at 200 mA in Tris-glycine buffer containing 20% methanol for 90 min at 4°C, and then put the membranes in 5% skim milk for block at 37°C for 1 h. The membranes were consistently incubated overnight at 4°C with diluted primary antibody that a rabbit polyclonal antibody against Nrf2 (Proteintech, Wuhan, China), HO-1 (Proteintech, Wuhan, China), p38MAPK (Abcam, Cambridge, UK), Bax (Servicebio Technology, Wuhan, China), Bcl-2 (Abcam, Cambridge, UK), Caspase-3 (CST, Trask Lane Danvers, MA).

Techniques: Sequencing

Journal: Poultry Science

Article Title: Marine algal polysaccharides alleviate aflatoxin B1-induced bursa of Fabricius injury by regulating redox and apoptotic signaling pathway in broilers

doi: 10.1016/j.psj.2020.10.050

Figure Lengend Snippet: The dilution ratio of antibodies in this study.

Article Snippet: Then, it was transferred to nitrocellulose membranes using a tank transfer at 200 mA in Tris-glycine buffer containing 20% methanol for 90 min at 4°C, and then put the membranes in 5% skim milk for block at 37°C for 1 h. The membranes were consistently incubated overnight at 4°C with diluted primary antibody that a rabbit polyclonal antibody against Nrf2 (Proteintech, Wuhan, China), HO-1 (Proteintech, Wuhan, China), p38MAPK (Abcam, Cambridge, UK), Bax (Servicebio Technology, Wuhan, China), Bcl-2 (Abcam, Cambridge, UK), Caspase-3 (CST, Trask Lane Danvers, MA).

Techniques:

Journal: Poultry Science

Article Title: Marine algal polysaccharides alleviate aflatoxin B1-induced bursa of Fabricius injury by regulating redox and apoptotic signaling pathway in broilers

doi: 10.1016/j.psj.2020.10.050

Figure Lengend Snippet: Effect of dietary aflatoxin B1 (AFB1) and marine algal polysaccharides (MAPs) on relative mRNA expression of antioxidant-related genes of the bursa of Fabricius in broilers. CON, control group, basal diet without addition of MAPs and AFB1; AFB1, basal diet with addition of 100 μg/kg AFB1; AFB1 + MAPs, basal diet with addition of 100 μg/kg and 2,500 mg/kg MAPs. (A) Relative mRNA expression of superoxide dismutase 1 (SOD1); (B) relative mRNA expression of superoxide dismutase 2 (SOD2); (C) relative mRNA expression of glutathione peroxidase 1 (GPx1); (D) relative mRNA expression of glutathione peroxidase 3 (GPx3); (E) relative mRNA expression of catalase 1 (CAT1); (F) relative mRNA expression of glutathione S-transferase theta 1 (GSTT1); (G) relative mRNA expression of glutathione S-transferase omega 1 (GSTO1); (H) relative mRNA expression of glutathione S-transferase alpha 3 (GSTA3); (I) relative mRNA expression of NF-E2-related factor 2 (Nrf2); (J) relative mRNA expression of heme oxygenase-1 (HO-1); (K) relative mRNA expression of p38 mitogen-activated protein (p38MAPK). All data were expressed as means ± SE (n = 6). ∗Means with an asterisk indicates a significant difference from control by Tukey's test (∗ P < 0.05; ∗∗ P < 0.01).

Article Snippet: Then, it was transferred to nitrocellulose membranes using a tank transfer at 200 mA in Tris-glycine buffer containing 20% methanol for 90 min at 4°C, and then put the membranes in 5% skim milk for block at 37°C for 1 h. The membranes were consistently incubated overnight at 4°C with diluted primary antibody that a rabbit polyclonal antibody against Nrf2 (Proteintech, Wuhan, China), HO-1 (Proteintech, Wuhan, China), p38MAPK (Abcam, Cambridge, UK), Bax (Servicebio Technology, Wuhan, China), Bcl-2 (Abcam, Cambridge, UK), Caspase-3 (CST, Trask Lane Danvers, MA).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: Marine algal polysaccharides alleviate aflatoxin B1-induced bursa of Fabricius injury by regulating redox and apoptotic signaling pathway in broilers

doi: 10.1016/j.psj.2020.10.050

Figure Lengend Snippet: Effect of dietary aflatoxin B1 (AFB1) and marine algal polysaccharides (MAPs) on protein expression levels of antioxidant and apoptosis-associated molecules of bursa of Fabricius in broilers. CON, control group, basal diet without addition of MAPs and AFB1; AFB1, basal diet with addition of 100 μg/kg AFB1; AFB1 + MAPs, basal diet with addition of 100 μg/kg and 2,500 mg/kg MAPs. (A) Protein expression levels of NF-E2-related factor 2 (Nrf2); (B) protein expression levels of heme oxygenase-1 (HO-1); (C) protein expression levels of p38 mitogen-activated protein (p38MAPK); (D) protein expression levels of Bcl-2-associated X (Bax); (E) protein expression levels of B-cell lymphoma-2 (Bcl-2); (F) protein expression levels of Caspase-3. All data were expressed as means ± SE (n = 6). ∗Means with an asterisk indicates a significant difference from control by Tukey's test (∗ P < 0.05; ∗∗ P < 0.01).

Article Snippet: Then, it was transferred to nitrocellulose membranes using a tank transfer at 200 mA in Tris-glycine buffer containing 20% methanol for 90 min at 4°C, and then put the membranes in 5% skim milk for block at 37°C for 1 h. The membranes were consistently incubated overnight at 4°C with diluted primary antibody that a rabbit polyclonal antibody against Nrf2 (Proteintech, Wuhan, China), HO-1 (Proteintech, Wuhan, China), p38MAPK (Abcam, Cambridge, UK), Bax (Servicebio Technology, Wuhan, China), Bcl-2 (Abcam, Cambridge, UK), Caspase-3 (CST, Trask Lane Danvers, MA).

Techniques: Expressing, Control

Journal: PLoS ONE

Article Title: Mechanisms and therapeutic implications of RTA 408, an activator of Nrf2, in subarachnoid hemorrhage–induced delayed cerebral vasospasm and secondary brain injury

doi: 10.1371/journal.pone.0240122

Figure Lengend Snippet: (A) Western blots of Nrf2, NF-κB and iNOS in the basilar artery. (B) SAH rats had significant reductions in Nrf2, NF-κB and iNOS. RTA 408(1mg/kg/day and 1.5 mg/kg/day) treatment reversed these changes, up-regulating Nrf2 and down-regulating NF-κB and iNOS. * p <0.05 and ** p <0.01 compared to controls. # p <0.05, and ## p <0.01 compared to SAH only.

Article Snippet: The next day the blots were incubated with following primary antibodies at the following dilutions: antibodies against rabbit polyclonal Nrf2 (Proteintech) 1:000, mouse monoclonal NF-κB (Cellsignaling) 1:500, mouse monoclonal iNOS (BD) 1:500, mouse monoclonal HO-1 (Ezno) 1:500, rabbit polyclonal NQO1 (Abcam) 1:1000, rabbit polyclonal cleaved caspase-3 (Cellsignaling) 1:500, and beta-actin (Sigma) 1:20000.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Mechanisms and therapeutic implications of RTA 408, an activator of Nrf2, in subarachnoid hemorrhage–induced delayed cerebral vasospasm and secondary brain injury

doi: 10.1371/journal.pone.0240122

Figure Lengend Snippet: (A) Western blots of Nrf2, HO-1, NQO1, cleaved caspase-3 and beta-action in the cortex, hippocampus, and dentate of rat brain. (B) Nrf2, HO-1 and NQO1 proteins were significantly reduced and cleaved caspase-3 was and the protein content of cleaved caspase-3 was significantly increased in the dentate gyrus untreated SAH rats. RTA-408 significantly increased Nrf2, HO-1 and NQO1 proteins and significantly decreased cleaved caspase 3 in the dentate gyrus of SAH rats.. There was no significant difference in these protein expressions in the cortexes of controls and untreated SAH rats and rats treated with RTA-408. * p <0.05 and ** p <0.01 compared to controls. # p <0.05, and ## p <0.01 compared to SAH only.

Article Snippet: The next day the blots were incubated with following primary antibodies at the following dilutions: antibodies against rabbit polyclonal Nrf2 (Proteintech) 1:000, mouse monoclonal NF-κB (Cellsignaling) 1:500, mouse monoclonal iNOS (BD) 1:500, mouse monoclonal HO-1 (Ezno) 1:500, rabbit polyclonal NQO1 (Abcam) 1:1000, rabbit polyclonal cleaved caspase-3 (Cellsignaling) 1:500, and beta-actin (Sigma) 1:20000.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Mechanisms and therapeutic implications of RTA 408, an activator of Nrf2, in subarachnoid hemorrhage–induced delayed cerebral vasospasm and secondary brain injury

doi: 10.1371/journal.pone.0240122

Figure Lengend Snippet: The RTA 408 achieves its anti-inflammatory, anti-oxidant and anti-apoptotic effects via Nrf2 and NF-κB signaling pathway.

Article Snippet: The next day the blots were incubated with following primary antibodies at the following dilutions: antibodies against rabbit polyclonal Nrf2 (Proteintech) 1:000, mouse monoclonal NF-κB (Cellsignaling) 1:500, mouse monoclonal iNOS (BD) 1:500, mouse monoclonal HO-1 (Ezno) 1:500, rabbit polyclonal NQO1 (Abcam) 1:1000, rabbit polyclonal cleaved caspase-3 (Cellsignaling) 1:500, and beta-actin (Sigma) 1:20000.

Techniques:

Journal: British Journal of Pharmacology

Article Title: 5‐fluorouracil causes endothelial cell senescence: potential protective role of glucagon‐like peptide 1

doi: 10.1111/bph.13725

Figure Lengend Snippet: 5FU induces senescence of endothelial cells. (A, B) Representative pictures and percentage of EA.hy926 cells stained for the senescence markers SA β‐gal (A) and p16INK4a (B) after no treatment (CTR) or exposure to 5FU. (C) Percentage of BrdU positive EA.hy926 cells in CTR condition and after incubation with 5FU. (D) Representative images of CTR and 5FU‐treated EA.hy926 cells stained for actin to visualize morphology and cytoplasm size. Nuclei are counterstained with DAPI. (E–G) Expression of VCAM1, ICAM1 and TYMP in CTR or 5FU‐treated cells. n = 7 for the experiments depicted in (A, B) and 5 for the other ones. * Statistically significant versus CTR (unpaired t‐test for A–C, Mann–Whitney test for E–G). Magnification of pictures is 400×, bars correspond to 50 μm.

Article Snippet: Immunocytochemistry for p16 INK4a was performed by using a primary rabbit polyclonal antibody against p16 INK4a (Proteintech, Rosemont, IL, USA) and Vectastain secondary antibody, HRP‐streptavidin and 3,3′‐diaminobenzidine (all from Vector Laboratories, Burlingame, CA, USA) to reveal the bound primary antibody.

Techniques: Staining, Incubation, Expressing, MANN-WHITNEY