rabbit polyclonal anti nrf1  (Proteintech)

Journal: Cell Death Discovery

Article Title: NRF1 coordinates mitochondrial adaptations to dampen intracellular ROS and inflammatory responses during ischemia reperfusion

doi: 10.1038/s41420-025-02461-5

Figure Lengend Snippet: A , B BMDMs were treated with OGD followed by different durations of reoxygenation. Nfe2l1 transcript was analyzed by qRT-PCR ( A ) and NRF1 proteins levels in cytosolic and nuclear fractions was analyzed by WB ( B ). C BMDMs were treated with OGD followed by different durations of reoxygenation +/− H 2 O 2 treatment during reoxygenation. NRF1 proteins levels in cytosolic and nuclear fractions was analyzed by WB. D BMDMs were treated with OGD followed by different durations of reoxygenation. NRF1 protein levels and mTORC1 signaling were assessed by WB. E BMDMs were subject to different durations of OGD followed by 1 h reoxygenation. NRF1 levels and mTORC1 signaling were assessed by WB. F BMDMs were subject to OGD/R +/− mTOR inhibitor (rapamycin, 400 nM) or S6K inhibitor (PF-4708671, 10 μM) during reoxygenation as indicated. NRF1 levels and mTORC1 signaling were assessed by WB. G Top , mouse model of ischemia and reperfusion injury induced acute kidney injury (IRI-AKI). Bottom , NRF1 MFI in F4/80 + CD11b + cells of kidneys subject to IRI-AKI or sham; quantitation and representative histogram are shown. Statistical test by one-way ANOVA and 2-tailed samples t-test ( n = 3–6 and representative of 3–6 independent experiments, n.s. no significance, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Single cell suspension was stained with antibodies for iNOS (CST,cat# 48866), CD206 (Biolegend,cat# 141717), CD11b (Biolegend,cat# 101207), F4/80 (BD Biosciences, cat# 567201), NRF1 (CST, cat# 8052S) and Alexa Fluor 647 anti-rabbit IgG (Abcam, cat# ab150075) followed by flow cytometry analysis.

Techniques: Quantitative RT-PCR, Quantitation Assay

Journal: Cell Death Discovery

Article Title: NRF1 coordinates mitochondrial adaptations to dampen intracellular ROS and inflammatory responses during ischemia reperfusion

doi: 10.1038/s41420-025-02461-5

Figure Lengend Snippet: A , B TOM20 staining of WT and NFE2L1 KO BMDMs and visualization by immunofluorescent confocal microscopy. Scale bar = 5 μm. Representative image in A; analysis and quantitation by Image J in ( B ). C WT and NFE2L1 KO BMDMs were subject to OGD/R +/− Tyrphostin A9 (10 μM) or Mdivi-1 (30 μM) during reoxygenation. DCFHA, MitoSox and TMRM staining was assessed by flow cytometry and MFI shown normalized to the untreated condition. Statistical test by two-way or one-way ANOVA ( n = 3–8 and representative of 3–8 independent experiments, n.s. no significance, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Single cell suspension was stained with antibodies for iNOS (CST,cat# 48866), CD206 (Biolegend,cat# 141717), CD11b (Biolegend,cat# 101207), F4/80 (BD Biosciences, cat# 567201), NRF1 (CST, cat# 8052S) and Alexa Fluor 647 anti-rabbit IgG (Abcam, cat# ab150075) followed by flow cytometry analysis.

Techniques: Staining, Confocal Microscopy, Quantitation Assay, Flow Cytometry

Journal: Cell Death Discovery

Article Title: NRF1 coordinates mitochondrial adaptations to dampen intracellular ROS and inflammatory responses during ischemia reperfusion

doi: 10.1038/s41420-025-02461-5

Figure Lengend Snippet: A Heatmap indicating relative expression levels of proteasome subunit genes differentially expressed between WT and NFE2L1 −/− BMDMs subject to OGD/R. B-C. WT and NFE2L1 −/− BMDMs were subject to OGD/R followed by analysis of poly-ubiquitinated proteins by WB ( B ). Intensityof Ubiquitin was normalized to α-Tubulin ( C ). D , E WT and NFE2L1 −/− BMDMs were subject to OGD/R followed by analysis of NRF1 in cytosolic and mitochondrial fractions by WB ( D ). Intensity of Ubiquitin was normalized to α-Tubulin in cytoplasm and VDAC1 in mitochondria ( E ). F , G WT and NFE2L1 −/− BMDMs were subject to OGD/R +/− Bafilomycin A1 (1 nM) treatment during reoxygenation. LC3 lipidation in mitochondrial fractions was assessed by WB ( F ). Data are presented as fold induction of LC3-II band intensity in the +BafA1 condition versus the -BafA1 condition for the corresponding OGD/R stimulation time point ( E ). H , I WT and NFE2L1 −/− BMDMs were subject to OGD/R +/− Bafilomycin A1 (1 nM) treatment during reoxygenation. LC3 puncta and mitochondria co-localization was assessed by immunofluorescent microscopy. Quantitation ( H ) and representative images ( I ) are shown. J , K WT and NFE2L1 −/− BMDMs were subject to OGD/R followed by analysis of the indicated mitophagy regulators by WB with ( J ) and qRT-PCR ( K ). L WT and NFE2L1 −/− BMDMs subject to OGD/R were analyzed for NRF1 binding to the promoters of the indicated genes by Cut&Tag-qPCR. Statistical test by two-way ANOVA ( n = 3 and representative of 3 independent experiments, n.s. no significance, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Single cell suspension was stained with antibodies for iNOS (CST,cat# 48866), CD206 (Biolegend,cat# 141717), CD11b (Biolegend,cat# 101207), F4/80 (BD Biosciences, cat# 567201), NRF1 (CST, cat# 8052S) and Alexa Fluor 647 anti-rabbit IgG (Abcam, cat# ab150075) followed by flow cytometry analysis.

Techniques: Expressing, Ubiquitin Proteomics, Microscopy, Quantitation Assay, Quantitative RT-PCR, Binding Assay

Journal: Cell Death Discovery

Article Title: NRF1 coordinates mitochondrial adaptations to dampen intracellular ROS and inflammatory responses during ischemia reperfusion

doi: 10.1038/s41420-025-02461-5

Figure Lengend Snippet: A WT and NFE2L1 −/− BMDMs were treated with OGD/R + /- LPS (10 ng/ml) or IL-4 (20 ng/ml), followed by analysis of Il6 and Arg1 gene expression by qRT-PCR. B WT and NFE2L1 −/− BMDMs were co-cultured with renal tubular epithelial cells during OGD/R, followed by analysis of Il6 and Arg1 gene expression by qRT-PCR. C-G. WT and mye-NFE2L1 KO mice were subject to IRI-AKI. C Kidney tissue sections visualized using hematoxylin-eosin (HE) staining. Blue arrow, normal renal tubules; black arrow, brush margin; red arrow, necrosis; green arrow, renal epithelial casts. D Kidney damage score: 0, no damage; 1, < 25%; 2, 25 to 50%; 3, 50 to 75%, 4, > 75%. E The proportion of iNOS + and CD206 + cells amongst kidney F4/80 + cells was determined by flow cytometry. F Serum levels of IL-6 as detected by ELISA. G Serum creatine (Jaffe’s method) and nitrogen (Jung’s method) were measured by spectrophotometer. Statistical test by one-way ANOVA and 2-tailed samples t-test ( n = 3–6 and representative of 3–6 independent experiments, n.s. no significance, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Single cell suspension was stained with antibodies for iNOS (CST,cat# 48866), CD206 (Biolegend,cat# 141717), CD11b (Biolegend,cat# 101207), F4/80 (BD Biosciences, cat# 567201), NRF1 (CST, cat# 8052S) and Alexa Fluor 647 anti-rabbit IgG (Abcam, cat# ab150075) followed by flow cytometry analysis.

Techniques: Gene Expression, Quantitative RT-PCR, Cell Culture, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Spectrophotometry

Journal: Cell Death Discovery

Article Title: NRF1 coordinates mitochondrial adaptations to dampen intracellular ROS and inflammatory responses during ischemia reperfusion

doi: 10.1038/s41420-025-02461-5

Figure Lengend Snippet: OGD/R triggers mitochondria to produce high levels of ROS, which activates the mTORC1 signaling pathway leading to the activation of NRF1. Activated NRF1 translocates to the nucleus, inducing genes regulating proteasome subunits to promote mitochondrial fission, which reduces mtROS production and facilitates mitophagy. Activated NRF1 also upregulates genes encoding mitophagy regulators to directly drive mitophagy. Thus, NRF1 coordinates mitochondrial adaptations that preserves mitochondrial integrity and reduces ROS levels and cellular stress. (Created with BioRender.com ).

Article Snippet: Single cell suspension was stained with antibodies for iNOS (CST,cat# 48866), CD206 (Biolegend,cat# 141717), CD11b (Biolegend,cat# 101207), F4/80 (BD Biosciences, cat# 567201), NRF1 (CST, cat# 8052S) and Alexa Fluor 647 anti-rabbit IgG (Abcam, cat# ab150075) followed by flow cytometry analysis.

Techniques: Activation Assay

Journal: Cell Genomics

Article Title: A highland-adaptation variant near MCUR1 reduces its transcription and attenuates erythrogenesis in Tibetans

doi: 10.1016/j.xgen.2025.100782

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-NRF1 , Proteintech , Cat#12482-1-AP; RRID: AB_2282876.

Techniques: Recombinant, Modification, Protease Inhibitor, Luminescence Assay, DNA Extraction, Multiplex Assay, Reporter Assay, Reverse Transcription, SYBR Green Assay, Electrophoresis, Mobility Shift, Transfection, Chromatin Immunoprecipitation, Mutagenesis, Enzyme-linked Immunosorbent Assay, Purification, Sequencing, RNA Sequencing, Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software