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Proteintech rabbit polyclonal anti nrf2 antibody
Reagents and instruments used in the study
Rabbit Polyclonal Anti Nrf2 Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti nrf2 antibody/product/Proteintech
Average 86 stars, based on 1 article reviews
rabbit polyclonal anti nrf2 antibody - by Bioz Stars, 2025-02
86/100 stars

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1) Product Images from "Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2"

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

Journal: Neural Regeneration Research

doi: 10.4103/NRR.NRR-D-23-01051

Reagents and instruments used in the study
Figure Legend Snippet: Reagents and instruments used in the study

Techniques Used: CCK-8 Assay, Saline, TUNEL Assay, DNA Extraction, Modification, Marker, Fluorescence, Microscopy, Imaging

NaHS reduces oxidative stress by suppressing Nrf2. (A) Representative images of DCFHDA staining in PC12 cells treated with Quin for 24 hours. The Quin group exhibited greater DCFH-DA (green) fluorescence than the Sham group. The Quin + NaHS group exhibited a decrease in DCFHDA (green) fluorescence compared with the Quin group, while an increase was observed in the Quin + NaHS + ML385 group. Scale bar: 50 μm. (B) DHE staining of PC12 cells following 24-hour Quin stimulation. Stronger DHE (red) fluorescence was observed in the Quin group compared with the Sham group. DHE (red) fluorescence was decreased in the Quin + NaHS group and increased in the Quin + NaHS + ML385 group compared with the Quin group. Scale bar: 50 μm. (C) DCFH-DA fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (D) DHE fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (E) DCFHDA fluorescence intensity in the striatum was quantified using a fluorescence microplate reader ( n = 5 per group). (F) Western blot detection of Nrf2 and HO-1 expression levels in PC12 cells in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). (G) Western blot detection of Nrf2 and HO-1 expression levels in the striatum of mice in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). The values are reported as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). DCFHDA: 2′,7′-Dichlorodihydrofluorescein diacetate; DHE: dihydroethidium; HO-1: heme oxygenase-1; Nrf2: nuclear factor erythroid 2-related factor 2; Quin: quinolinic acid.
Figure Legend Snippet: NaHS reduces oxidative stress by suppressing Nrf2. (A) Representative images of DCFHDA staining in PC12 cells treated with Quin for 24 hours. The Quin group exhibited greater DCFH-DA (green) fluorescence than the Sham group. The Quin + NaHS group exhibited a decrease in DCFHDA (green) fluorescence compared with the Quin group, while an increase was observed in the Quin + NaHS + ML385 group. Scale bar: 50 μm. (B) DHE staining of PC12 cells following 24-hour Quin stimulation. Stronger DHE (red) fluorescence was observed in the Quin group compared with the Sham group. DHE (red) fluorescence was decreased in the Quin + NaHS group and increased in the Quin + NaHS + ML385 group compared with the Quin group. Scale bar: 50 μm. (C) DCFH-DA fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (D) DHE fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (E) DCFHDA fluorescence intensity in the striatum was quantified using a fluorescence microplate reader ( n = 5 per group). (F) Western blot detection of Nrf2 and HO-1 expression levels in PC12 cells in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). (G) Western blot detection of Nrf2 and HO-1 expression levels in the striatum of mice in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). The values are reported as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). DCFHDA: 2′,7′-Dichlorodihydrofluorescein diacetate; DHE: dihydroethidium; HO-1: heme oxygenase-1; Nrf2: nuclear factor erythroid 2-related factor 2; Quin: quinolinic acid.

Techniques Used: Staining, Fluorescence, Western Blot, Expressing

NaHS regulates mitochondrial function in Quin-stimulated PC12 cells by suppressing Nrf2. (A) Mitochondria stained with MitoTracker (red) in Quin-stimulated PC12 cells treated with NaHS and ML385. The Quin group exhibited severe mitochondrial fragmentation and a decrease in mitochondrial fluorescence intensity compared with the Sham group, whereas the addition of NaHS resulted in an increase in mitochondrial fluorescence intensity. However, this effect was reversed by the addition of ML385. Scale bar: 10 μm. (B) Quantification of mitochondrial fluorescence intensity ( n = 10 per group). (C) Representative micrographs of MMP in Quin-stimulated PC12 cells, as detected via JC-1. The transition from green to red fluorescence indicates mitochondrial polarization, which was restored to normal levels by NaHS treatment. In contrast, ML385 treatment induced a shift from green to red fluorescence, indicating substantial mitochondrial depolarization. Scale bar: 50 μm. (D) Quantification of mitochondrial MMP ( n = 15 per group). (E) Quantification of mitochondrial DNA (mtDNA, NADH5) versus nuclear DNA (GAPDH) as detected by real-time polymerase chain reaction in Quin-stimulated PC12 cells treated with NaHS and ML385. (F) Western blot detection of TOMM20 expression levels in PC12 cells. Quantification of TOMM20 expression ( n = 3 per group). The values are reported as mean ± SD. ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; MMP: mitochondrial membrane potential; Quin: quinolinic acid; TOMM20: translocase of outer mitochondrial membrane 20.
Figure Legend Snippet: NaHS regulates mitochondrial function in Quin-stimulated PC12 cells by suppressing Nrf2. (A) Mitochondria stained with MitoTracker (red) in Quin-stimulated PC12 cells treated with NaHS and ML385. The Quin group exhibited severe mitochondrial fragmentation and a decrease in mitochondrial fluorescence intensity compared with the Sham group, whereas the addition of NaHS resulted in an increase in mitochondrial fluorescence intensity. However, this effect was reversed by the addition of ML385. Scale bar: 10 μm. (B) Quantification of mitochondrial fluorescence intensity ( n = 10 per group). (C) Representative micrographs of MMP in Quin-stimulated PC12 cells, as detected via JC-1. The transition from green to red fluorescence indicates mitochondrial polarization, which was restored to normal levels by NaHS treatment. In contrast, ML385 treatment induced a shift from green to red fluorescence, indicating substantial mitochondrial depolarization. Scale bar: 50 μm. (D) Quantification of mitochondrial MMP ( n = 15 per group). (E) Quantification of mitochondrial DNA (mtDNA, NADH5) versus nuclear DNA (GAPDH) as detected by real-time polymerase chain reaction in Quin-stimulated PC12 cells treated with NaHS and ML385. (F) Western blot detection of TOMM20 expression levels in PC12 cells. Quantification of TOMM20 expression ( n = 3 per group). The values are reported as mean ± SD. ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; MMP: mitochondrial membrane potential; Quin: quinolinic acid; TOMM20: translocase of outer mitochondrial membrane 20.

Techniques Used: Staining, Fluorescence, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Membrane



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Reagents and instruments used in the study

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: Reagents and instruments used in the study

Article Snippet: Rabbit polyclonal anti-NRF2 antibody , Proteintech Group , 16396-1-AP (RRID: AB_2782956).

Techniques: CCK-8 Assay, Saline, TUNEL Assay, DNA Extraction, Modification, Marker, Fluorescence, Microscopy, Imaging

NaHS reduces oxidative stress by suppressing Nrf2. (A) Representative images of DCFHDA staining in PC12 cells treated with Quin for 24 hours. The Quin group exhibited greater DCFH-DA (green) fluorescence than the Sham group. The Quin + NaHS group exhibited a decrease in DCFHDA (green) fluorescence compared with the Quin group, while an increase was observed in the Quin + NaHS + ML385 group. Scale bar: 50 μm. (B) DHE staining of PC12 cells following 24-hour Quin stimulation. Stronger DHE (red) fluorescence was observed in the Quin group compared with the Sham group. DHE (red) fluorescence was decreased in the Quin + NaHS group and increased in the Quin + NaHS + ML385 group compared with the Quin group. Scale bar: 50 μm. (C) DCFH-DA fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (D) DHE fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (E) DCFHDA fluorescence intensity in the striatum was quantified using a fluorescence microplate reader ( n = 5 per group). (F) Western blot detection of Nrf2 and HO-1 expression levels in PC12 cells in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). (G) Western blot detection of Nrf2 and HO-1 expression levels in the striatum of mice in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). The values are reported as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). DCFHDA: 2′,7′-Dichlorodihydrofluorescein diacetate; DHE: dihydroethidium; HO-1: heme oxygenase-1; Nrf2: nuclear factor erythroid 2-related factor 2; Quin: quinolinic acid.

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: NaHS reduces oxidative stress by suppressing Nrf2. (A) Representative images of DCFHDA staining in PC12 cells treated with Quin for 24 hours. The Quin group exhibited greater DCFH-DA (green) fluorescence than the Sham group. The Quin + NaHS group exhibited a decrease in DCFHDA (green) fluorescence compared with the Quin group, while an increase was observed in the Quin + NaHS + ML385 group. Scale bar: 50 μm. (B) DHE staining of PC12 cells following 24-hour Quin stimulation. Stronger DHE (red) fluorescence was observed in the Quin group compared with the Sham group. DHE (red) fluorescence was decreased in the Quin + NaHS group and increased in the Quin + NaHS + ML385 group compared with the Quin group. Scale bar: 50 μm. (C) DCFH-DA fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (D) DHE fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (E) DCFHDA fluorescence intensity in the striatum was quantified using a fluorescence microplate reader ( n = 5 per group). (F) Western blot detection of Nrf2 and HO-1 expression levels in PC12 cells in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). (G) Western blot detection of Nrf2 and HO-1 expression levels in the striatum of mice in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). The values are reported as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). DCFHDA: 2′,7′-Dichlorodihydrofluorescein diacetate; DHE: dihydroethidium; HO-1: heme oxygenase-1; Nrf2: nuclear factor erythroid 2-related factor 2; Quin: quinolinic acid.

Article Snippet: Rabbit polyclonal anti-NRF2 antibody , Proteintech Group , 16396-1-AP (RRID: AB_2782956).

Techniques: Staining, Fluorescence, Western Blot, Expressing

NaHS regulates mitochondrial function in Quin-stimulated PC12 cells by suppressing Nrf2. (A) Mitochondria stained with MitoTracker (red) in Quin-stimulated PC12 cells treated with NaHS and ML385. The Quin group exhibited severe mitochondrial fragmentation and a decrease in mitochondrial fluorescence intensity compared with the Sham group, whereas the addition of NaHS resulted in an increase in mitochondrial fluorescence intensity. However, this effect was reversed by the addition of ML385. Scale bar: 10 μm. (B) Quantification of mitochondrial fluorescence intensity ( n = 10 per group). (C) Representative micrographs of MMP in Quin-stimulated PC12 cells, as detected via JC-1. The transition from green to red fluorescence indicates mitochondrial polarization, which was restored to normal levels by NaHS treatment. In contrast, ML385 treatment induced a shift from green to red fluorescence, indicating substantial mitochondrial depolarization. Scale bar: 50 μm. (D) Quantification of mitochondrial MMP ( n = 15 per group). (E) Quantification of mitochondrial DNA (mtDNA, NADH5) versus nuclear DNA (GAPDH) as detected by real-time polymerase chain reaction in Quin-stimulated PC12 cells treated with NaHS and ML385. (F) Western blot detection of TOMM20 expression levels in PC12 cells. Quantification of TOMM20 expression ( n = 3 per group). The values are reported as mean ± SD. ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; MMP: mitochondrial membrane potential; Quin: quinolinic acid; TOMM20: translocase of outer mitochondrial membrane 20.

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: NaHS regulates mitochondrial function in Quin-stimulated PC12 cells by suppressing Nrf2. (A) Mitochondria stained with MitoTracker (red) in Quin-stimulated PC12 cells treated with NaHS and ML385. The Quin group exhibited severe mitochondrial fragmentation and a decrease in mitochondrial fluorescence intensity compared with the Sham group, whereas the addition of NaHS resulted in an increase in mitochondrial fluorescence intensity. However, this effect was reversed by the addition of ML385. Scale bar: 10 μm. (B) Quantification of mitochondrial fluorescence intensity ( n = 10 per group). (C) Representative micrographs of MMP in Quin-stimulated PC12 cells, as detected via JC-1. The transition from green to red fluorescence indicates mitochondrial polarization, which was restored to normal levels by NaHS treatment. In contrast, ML385 treatment induced a shift from green to red fluorescence, indicating substantial mitochondrial depolarization. Scale bar: 50 μm. (D) Quantification of mitochondrial MMP ( n = 15 per group). (E) Quantification of mitochondrial DNA (mtDNA, NADH5) versus nuclear DNA (GAPDH) as detected by real-time polymerase chain reaction in Quin-stimulated PC12 cells treated with NaHS and ML385. (F) Western blot detection of TOMM20 expression levels in PC12 cells. Quantification of TOMM20 expression ( n = 3 per group). The values are reported as mean ± SD. ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; MMP: mitochondrial membrane potential; Quin: quinolinic acid; TOMM20: translocase of outer mitochondrial membrane 20.

Article Snippet: Rabbit polyclonal anti-NRF2 antibody , Proteintech Group , 16396-1-AP (RRID: AB_2782956).

Techniques: Staining, Fluorescence, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Membrane

Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Western Blot, Immunohistochemistry

The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Quantitative RT-PCR, Western Blot

The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques: Quantitative RT-PCR, Western Blot, Immunohistochemistry, Activity Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining

Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques:

Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Western Blot, Immunohistochemistry

The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Quantitative RT-PCR, Western Blot

The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques: Quantitative RT-PCR, Western Blot, Immunohistochemistry, Activity Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining

Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques:

Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Western Blot, Immunohistochemistry

The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Quantitative RT-PCR, Western Blot

The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques: Quantitative RT-PCR, Western Blot, Immunohistochemistry, Activity Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining

Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Article Snippet: Application of primary antibodies consisted of rabbit anti-Nrf2 polyclonal antibody (rat, PA5-27882, 1:500, ThermoFisher, USA), rabbit anti-p-Nrf2 polyclonal antibody (rat, PA5-102838, 1:200, ThermoFisher), and rabbit anti-HO-1 polyclonal antibody (rat, PA5-77834, 1:1000, ThermoFisher), and incubation at 4°C throughout the night was applied to the sections.

Techniques:

Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Impact of NEAT1 on the levels of factors related to the Nrf2/ARE/HO-1 signaling axis. Note: A Intersection results of lncRNA NEAT1 target factors predicted by starBase v2.0, RNAInter, and AnnoLnc2 databases; B Changes in the expression levels of NR3C1, RBFOX2, TAF15, STAG1, NFE2L2 (Nrf2), and EZH2 detected by RT-qPCR after silencing NEAT1 in rat hippocampal tissue; C EMSA experiment using ARE probe and lentivirus transfection in neuronal nuclear extracts of the control group and NEAT1 silenced group, with Supershift assay using anti-Nrf2 antibody. The complexes induced by NEAT1 silencing are marked by arrows, and asterisks denote the supershifted complexes; D mRNA levels of HO-1 and Nrf2 in hippocampal tissue of rats from each group detected by RT-qPCR; E Protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in hippocampal tissue of rats in each group examined by Western blot; F Immunohistochemistry analysis of the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissue, with a scale bar of 50 μm. Each group consisted of 8 rats. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Western Blot, Immunohistochemistry

The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on oxidative stress and apoptosis in primary hippocampal neurons via the NEAT1/Nrf2/ARE/HO-1 axis. Note: A Detection of ROS activity in neuronal cells of rats in each group; B Measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group using ELISA; C Assessment of apoptosis in neuronal cells of rats in each group using flow cytometry; D Quantification of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group through RT-qPCR; E Evaluation of protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary hippocampal neurons of rats in each group by Western blot; F RT-qPCR analysis of NEAT1, HO-1, and Nrf2 mRNA levels in primary hippocampal neurons of rats in each group; G Western blot assessment of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 protein levels in primary hippocampal neurons of rats in each group; H Detection of ROS activity in neuronal cells of rats in each group; I ELISA measurement of MDA, SOD, and GSH levels in neuronal cells of rats in each group; J Flow cytometry analysis of apoptosis in neuronal cells of rats in each group. All cell experiments were repeated three times. Indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Quantitative RT-PCR, Western Blot

The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: The impact of sevoflurane on hippocampal neuronal damage and cognitive dysfunction in rats through the NEAT1/Nrf2/ARE/HO-1 signaling pathway. Note: A RT-qPCR was used to measure the mRNA levels of NEAT1, HO-1, and Nrf2 in primary rat hippocampal neurons in each group; B Western blot was performed to assess the protein levels of HO-1, NQO1, catalase, MnSOD, Nrf2, and p-Nrf2 in primary rat hippocampal neurons in each group; C Immunohistochemistry was conducted to determine the number of positive cells for HO-1, Nrf2, and p-Nrf2 in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; D - E MWM test was utilized to evaluate the memory capacity of rats in each group, including escape latency (D) and number of platform crossings (E); F Measurement of ROS activity in hippocampal tissues of each group of rats; G ELISA was employed to analyze the levels of MDA, SOD, and GSH in hippocampal tissues of each group of rats; H TUNEL assay was used to assess the neuronal apoptosis in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm; I H&E staining was performed to evaluate the extent of damage in the CA1 region of rat hippocampal tissues in each group, with a scale bar of 50 μm. Each group consisted of 8 rats. Data at different time points were analyzed using repeated measures ANOVA, other indicators between two groups were compared using independent sample t-tests, and multiple group comparisons were analyzed using one-way ANOVA followed by Tukey's post hoc test. The lines connecting the two groups in the figure indicate a significant difference at P < 0.05

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques: Quantitative RT-PCR, Western Blot, Immunohistochemistry, Activity Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining

Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Journal: Cell Biology and Toxicology

Article Title: Mechanistic insights into sevoflurane-induced hippocampal neuronal damage and cognitive dysfunction through the NEAT1/Nrf2 signaling axis in aged rats

doi: 10.1007/s10565-024-09964-4

Figure Lengend Snippet: Molecular mechanism of sevoflurane regulation of the lncRNA NEAT1/Nrf2 signaling axis on hippocampal neuronal damage and cognitive dysfunction in aged rats

Article Snippet: The loading of 30 µg of cell lysate onto SDS-PAGE was tracked by the transfer onto a nitrocellulose (NC) membrane, followed by blocking with 5% skim milk in TBST for a duration of 1.5 h. Primary antibodies used included rabbit polyclonal anti-Nrf2 (PA5-27882, 1:1000), rabbit polyclonal anti-phosphorylated Nrf2 (PA5-102838, 1:1000), rabbit polyclonal anti-HO-1 (PA5-77834, 1:1000), rabbit polyclonal anti-NQO1 (PA5-21290, 1:1000), rabbit polyclonal anti-catalase (PA5-29183, 1:1000), and rabbit polyclonal anti-MnSOD (PA5-30604, 1:1000) from ThermoFisher, USA.

Techniques: