Journal: bioRxiv

Article Title: Temporal Coordination of the Transcription Factor Response to H 2 O 2 stress

doi: 10.1101/2023.03.07.531593

Figure Lengend Snippet: (A) Uniform manifold approximation and projection (UMAP) plot of cells treated with PBS, 50μM and 75μM of H 2 O 2 after unsupervised clustering (n ≥ 10,000). Colors for cells based on the six clusters obtained. (B) UMAP of the same cells in A but colored based on sample. (C) UMAPs of cells from A colored by deviation scores for FOXO1 (left), RelA (middle) and HSF1 motifs (right) (D) UMAPs of cells from A showing deviation scores for p53 (left), Jun(middle) and NRF2 motifs (right) (E) Density colored scatter plots of log nuclear p53 (x-axis) and nuclear fraction of RelA (y-axis) at indicated levels of H 2 O 2 treatment for 5 hours (F) Percentage of cells activating both RelA and p53 (Both), RelA only, p53 only or neither for all concentrations of H 2 O 2 (G) Density colored scatter plots of log nuclear p53 (x-axis) and nuclear fraction of NFAT1 (y-axis) at indicated levels of H 2 O 2 treatment for 5 hours (H) Percentage of cells activating both NFAT1 and p53 (Both), NFAT1 only, p53 only or neither for all concentrations of H 2 O 2 . (I) Density colored scatter plots of log nuclear NRF2 (x-axis) and nuclear fraction of FOXO1(y-axis) at indicated levels of H 2 O 2 treatment for 5 hours (J) Percentage of cells activating both FOXO1 and NRF2 (Both), FOXO1 only, NRF2 only or neither for all concentrations of H 2 O 2 . (K) Density colored scatter plots of log nuclear JUN (x-axis) and nuclear fraction of FOXO1 (y-axis) at indicated levels of H 2 O 2 treatment for 5 hours (L) Percentage of cells activating both FOXO1 and JUN (Both), FOXO1 only, JUN only or neither for all concentrations of H 2 O 2 .

Article Snippet: Primary antibodies used: Anti-FOXO1 (C29H4) from Cell Signaling Cat# 2880S (1:500), Anti-p53 (DO-1) from Santa Cruz Cat# sc-126 (1:500), Anti-Sulfiredoxin from Santa Cruz Cat# sc-166786 (1:100), Recombinant Anti-Peroxiredoxin 1/PAG antibody [EPR5433] (ab109498) (1:1000), NRF2 (D1Z9C) XP Rabbit mAb #12721 (1:500), NFAT1 (D43B1) XP Rabbit mAb #5861 (1:300), NF-kB p65 (D14E12) XP Rabbit mAb #8242 (1:400), Anti-HSF1 antibody 10H8 from Stress Marq Biosciences (1:200), c-Fos (9F6) Rabbit mAb # 2250 (1:1000), and c-Jun (60A8) Rabbit mAb #9165 (1:400) from Cell Signaling.

Techniques:

Journal: bioRxiv

Article Title: Temporal Coordination of the Transcription Factor Response to H 2 O 2 stress

doi: 10.1101/2023.03.07.531593

Figure Lengend Snippet: (A) GSEA of p53 target genes in J14 + H 2 O 2 samples as compared to SRXN1-OE + H 2 O 2 samples (B) GSEA of NRF2 target genes in J14 + H 2 O 2 samples as compared to SRXN1-OE + H 2 O 2 samples. (C) Wald statistic and (D) log 2 fold changes of p53 target genes in H 2 O 2 treated cells vs. PBS controls, H 2 O 2 + J14 treated cells vs J14 treated controls and H 2 O 2 treated SRXN1-OE cells vs. SRXN1-OE PBS controls. (E) Wald statistic and (F) log 2 fold changes of NRF2 target genes in H 2 O 2 treated cells vs. PBS controls, H 2 O 2 + J14 treated cells vs J14 treated controls and H 2 O 2 treated SRXN1-OE cells vs. SRXN1-OE PBS controls. H 2 O 2 concentration is 50μM, J14 20μM.

Article Snippet: Primary antibodies used: Anti-FOXO1 (C29H4) from Cell Signaling Cat# 2880S (1:500), Anti-p53 (DO-1) from Santa Cruz Cat# sc-126 (1:500), Anti-Sulfiredoxin from Santa Cruz Cat# sc-166786 (1:100), Recombinant Anti-Peroxiredoxin 1/PAG antibody [EPR5433] (ab109498) (1:1000), NRF2 (D1Z9C) XP Rabbit mAb #12721 (1:500), NFAT1 (D43B1) XP Rabbit mAb #5861 (1:300), NF-kB p65 (D14E12) XP Rabbit mAb #8242 (1:400), Anti-HSF1 antibody 10H8 from Stress Marq Biosciences (1:200), c-Fos (9F6) Rabbit mAb # 2250 (1:1000), and c-Jun (60A8) Rabbit mAb #9165 (1:400) from Cell Signaling.

Techniques: Concentration Assay

Journal: Biomedicines

Article Title: Cadmium-Induced Kidney Injury in Mice Is Counteracted by a Flavonoid-Rich Extract of Bergamot Juice, Alone or in Association with Curcumin and Resveratrol, via the Enhancement of Different Defense Mechanisms

doi: 10.3390/biomedicines9121797

Figure Lengend Snippet: Oligonucleotide primers used for the quantitative Real-time PCR analyses.

Article Snippet: Primary antibodies IL-1β (1:250, Santa Cruz Biotechnology, Dallas, TX, USA) and Nrf2 (1:150, St. John’s Laboratory, London, UK) were incubated overnight at 4 °C in a moisturized chamber.

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Biomedicines

Article Title: Cadmium-Induced Kidney Injury in Mice Is Counteracted by a Flavonoid-Rich Extract of Bergamot Juice, Alone or in Association with Curcumin and Resveratrol, via the Enhancement of Different Defense Mechanisms

doi: 10.3390/biomedicines9121797

Figure Lengend Snippet: Real-time PCR analysis of Nrf2 ( A ), Nqo1 ( B ) and Hmox1 ( C ). *** p < 0.001 vs. control mice; §§§ p < 0.001 vs. CdCl 2 -treated mice.

Article Snippet: Primary antibodies IL-1β (1:250, Santa Cruz Biotechnology, Dallas, TX, USA) and Nrf2 (1:150, St. John’s Laboratory, London, UK) were incubated overnight at 4 °C in a moisturized chamber.

Techniques: Real-time Polymerase Chain Reaction, Control

Journal: Biomedicines

Article Title: Cadmium-Induced Kidney Injury in Mice Is Counteracted by a Flavonoid-Rich Extract of Bergamot Juice, Alone or in Association with Curcumin and Resveratrol, via the Enhancement of Different Defense Mechanisms

doi: 10.3390/biomedicines9121797

Figure Lengend Snippet: Immunohistochemical localization of Nrf2 in the kidneys. ( A ) In all control groups, Nrf2 immunoreactivity is particularly strong in the tubular wall (arrow). ( B ) In CdCl 2 plus vehicle-treated mice, no Nrf2 immunoreactivity is present. ( C – E ) In mice treated with CdCl 2 plus both doses of Cur and with CdCl 2 plus Re, Nrf2 shows a moderate positivity in some tubules (arrow). ( F ) In mice challenged with CdCl 2 plus BJe at the lower dose, Nrf2 immunoreactivity was lower (arrow) when compared to Cur and Re. ( G – I ) Mice treated with CdCl 2 plus BJe at 40 mg/kg and with CdCl 2 plus both associations: Nrf2 immunoreactivity is high (arrow), similar to controls. ( J ) Morphometric results for Nrf2 expression. Data are expressed in optical units/unit area (OU/UA) (from 0 = black to 255 = white). * p < 0.05 vs. control; § p < 0.05 vs. CdCl 2 plus vehicle. Scale bar: 50 µm.

Article Snippet: Primary antibodies IL-1β (1:250, Santa Cruz Biotechnology, Dallas, TX, USA) and Nrf2 (1:150, St. John’s Laboratory, London, UK) were incubated overnight at 4 °C in a moisturized chamber.

Techniques: Immunohistochemical staining, Control, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Protective effects of Rosavin on bleomycin-induced pulmonary fibrosis via suppressing fibrotic and inflammatory signaling pathways in mice.

doi: 10.1016/j.biopha.2019.108870

Figure Lengend Snippet: Fig. 8. The effect of Rosavin on the expression of TGF-β1, α-SMA, Nrf2 and NF-κB p65 of BIM-induced PF in mice. Values are the mean ± SD (n = 3); nsP > 0.05, ##P < 0.01 vs. control group. *P < 0.05, **P < 0.01 vs. Bleomycin control group.

Article Snippet: Rabbit anti-TGF-β1 antibody (21898-1-AP) was purchased from Proteintech (Wuhan, China), and mouse anti-α-SMA antibody (BM0002), rabbit anti-NF-Kb p65 antibody, rabbit anti-Nrf2 antibody were all obtained from Wuhan Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Expressing, Control

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Tempol relieves lung injury in a rat model of chronic intermittent hypoxia via suppression of inflammation and oxidative stress

doi: 10.22038/ijbms.2018.31716.7714

Figure Lengend Snippet: Tempol further promoted intermittent hypoxia-induced activation of the Nrf2/HO-1 signaling pathway. (A) The level of HO-1 in lung tissues was detected by a commercial kit. (B) The protein levels of HO-1, cytoplasmic Nrf2, and nuclear Nrf2 in lung tissues were measured by Western blot assay. GAPDH was used as a loading control. (C)& (D) The gray-scale value of the bands was quantitatively analyzed. (E) DNA binding activity of Nrf2 in lung tissues was determined by EMSA. (F) Quantitative analysis of Nrf2 DNA binding activity. The experimental data were expressed as mean±SD (n=6). * P< 0.05, *** P< 0.001, versus the NA group. ### P< 0.001, versus the IH group. ns, no significance, versus the NA group

Article Snippet: Whereafter the membranes were blocked with 5% skimmed milk for 1 hr and incubated with corresponding primary antibodies against COX2 (1:700, Proteintech, USA), iNOS (1:7500, Proteintech, USA), IκBα (1:1000, Cell Signaling Technology, USA), p- IκBα (1:1000, Cell Signaling Technology, USA), NF-κB (1:1000, Proteintech, USA), HO-1 (1:500, Boster, China), Nrf2 (1:500 Boster, China), GAPDH (1:5000, Bioss, China), and Histone H3 (1:1000, Proteintech, USA) at 4 ° C overnight.

Techniques: Activation Assay, Western Blot, Control, Binding Assay, Activity Assay