Journal: Basic and clinical andrology

Article Title: The protective effects of active ingredients from acrorus tatarinowii on sperm and their molecular mechanisms.

doi: 10.1186/s12610-024-00247-w

Figure Lengend Snippet: Fig. 3 Acrorus tatarinowii controls the BCL2-related pathway in the oligoasthenozoospermia rat. a-j qPCR analysis of mRNA expression of TNF-α, MAPK14, GSPT1, KDR, HMOX1, GSTM1, ESR1, CYP1B1, CYP1A1, and BCL2 in rat testicular tissues. K Western blot detection of protein expression of each gene. n = 3,vs.control,*P < 0.05,**P < 0.01,***P < 0.001 vs.Oligoasthenozoospermia,&P < 0.05,&&P < 0.01, &&&P < 0.001, ns, no significance

Article Snippet: Subsequently, proteins were transferred onto membranes and incubated with primary antibodies against TNF-α (CST), MAPK14 (CST), KDR (CST), HMOX1 (CST), GSTP1 (CST), GSTM1 (Proteintech), ESR1 (Proteintech), CYP1B1 (Proteintech), CYP1A1 (Proteintech), and BCL2 (Abcam).

Techniques: Expressing, Western Blot, Control

Journal: Basic and clinical andrology

Article Title: The protective effects of active ingredients from acrorus tatarinowii on sperm and their molecular mechanisms.

doi: 10.1186/s12610-024-00247-w

Figure Lengend Snippet: Fig. 3 Acrorus tatarinowii controls the BCL2-related pathway in the oligoasthenozoospermia rat. a-j qPCR analysis of mRNA expression of TNF-α, MAPK14, GSPT1, KDR, HMOX1, GSTM1, ESR1, CYP1B1, CYP1A1, and BCL2 in rat testicular tissues. K Western blot detection of protein expression of each gene. n = 3,vs.control,*P < 0.05,**P < 0.01,***P < 0.001 vs.Oligoasthenozoospermia,&P < 0.05,&&P < 0.01, &&&P < 0.001, ns, no significance

Article Snippet: Subsequently, proteins were transferred onto membranes and incubated with primary antibodies against TNF-α (CST), MAPK14 (CST), KDR (CST), HMOX1 (CST), GSTP1 (CST), GSTM1 (Proteintech), ESR1 (Proteintech), CYP1B1 (Proteintech), CYP1A1 (Proteintech), and BCL2 (Abcam).

Techniques: Expressing, Western Blot, Control

Journal: Journal of Toxicologic Pathology

Article Title: Thioacetamide-induced Hepatocellular Necrosis Is Attenuated in Diet-induced Obese Mice

doi: 10.1293/tox.26.175

Figure Lengend Snippet: Western blots of hepatic drug-metabolizing enzymes in the livers of vehicle-treated mice. The number of mice fed the high-fat diet (HFD) or normal rodent diet (ND) was 4. A: Western blots of CYP2E1, glutathione S -transferases (GST) A1, GSTA2, GSTA3, GSTA4, GSTP1 and GSTM1 of HFD mice and ND mice are shown. B: Expression of drug-metabolizing enzymes as quantified by densitometry is shown. Black columns show the relative expression ratios of HFD mice to ND mice. *, ** p<0.05 or p<0.01 versus the expression in mice fed the normal rodent diet, respectively (Aspin-Welch t-test).

Article Snippet: To detect the drug-metabolizing enzyme expression, antibodies against CYP2E1 (Chemicon International, Temecula, CA, USA), GSTA1, GSTA2, GSTA3, GSTA4, GSTP1 and GSTM1 (Proteintech Group, Inc., Chicago, IL, USA) were used.

Techniques: Western Blot, Expressing

Journal: Journal of Toxicologic Pathology

Article Title: Thioacetamide-induced Hepatocellular Necrosis Is Attenuated in Diet-induced Obese Mice

doi: 10.1293/tox.26.175

Figure Lengend Snippet: Western blots of hepatic drug-metabolizing enzymes in the livers of vehicle-treated mice. The number of mice fed the high-fat diet (HFD) or normal rodent diet (ND) was 4. A: Western blots of CYP2E1, glutathione S -transferases (GST) A1, GSTA2, GSTA3, GSTA4, GSTP1 and GSTM1 of HFD mice and ND mice are shown. B: Expression of drug-metabolizing enzymes as quantified by densitometry is shown. Black columns show the relative expression ratios of HFD mice to ND mice. *, ** p<0.05 or p<0.01 versus the expression in mice fed the normal rodent diet, respectively (Aspin-Welch t-test).

Article Snippet: To detect the drug-metabolizing enzyme expression, antibodies against CYP2E1 (Chemicon International, Temecula, CA, USA), GSTA1, GSTA2, GSTA3, GSTA4, GSTP1 and GSTM1 (Proteintech Group, Inc., Chicago, IL, USA) were used.

Techniques: Western Blot, Expressing

Journal: The FASEB Journal

Article Title: Gstm1/Gstt1 is essential for reducing cisplatin ototoxicity in CBA/CaJ mice

doi: 10.1096/fj.202200324r

Figure Lengend Snippet: FIGURE 1 Gstm1 and Gstt1 mRNA and protein expression in the cochleae of cisplatin-treated CBA/CaJ mice. (A) Schematic strategy for cisplatin treatment in CBA/CaJ mice. (B) ABR hearing thresholds in CBA/CaJ mice before and after cisplatin treatment. n = 6. (C) mRNA expression levels of Gstm1 and Gstt1 in the cochleae of CBA/CaJ mice before and after cisplatin treatment. n = 3. (D) Protein expression levels of Gstm1 and Gstt1 in the cochleae of CBA/CaJ mice before and after cisplatin treatment. (E) Quantitative statistics of relative protein expression of Gstm1 and Gstt1 in the cochleae of CBA/CaJ mice before and after cisplatin treatment. n = 5. *p < .05, **p < .01, ***p < .001. Data are expressed as the mean ± SEM. ABR, auditory brainstem response.

Article Snippet: The following primary antibodies were used: Gstm1 (rabbit polyclonal, 1:4000, Proteintech, 12412- 1- AP), Gstt1 (rabbit polyclonal, 1:2000, Proteintech, 15838- 1- AP). p- P38 (rabbit polyclonal, 1:1000, Cell Signaling Technology, 9211S), p- JNK (rabbit polyclonal, 1:1000, Cell Signaling Technology, 4668T), p- ERK (rabbit polyclonal, 1:2000, Cell Signaling Technology, 4370T), and pAkt (rabbit polyclonal, 1:1000, Cell Signaling Technology, 13038S).

Techniques: Expressing

Journal: The FASEB Journal

Article Title: Gstm1/Gstt1 is essential for reducing cisplatin ototoxicity in CBA/CaJ mice

doi: 10.1096/fj.202200324r

Figure Lengend Snippet: FIGURE 2 Generation of Gstm1−/− and Gstt1−/− mice. (A) Schematic strategy of Gstm1 knockout in mice. (B) Schematic strategy of Gstt1 knockout in mice. (C) mRNA expression levels of Gstm1 and Gstt1 in the cochleae of WT, Gstm1−/−, and Gstt1−/− mice. n = 3, ***p < .001. (D) Protein expression levels of Gstm1 and Gstt1 in the cochleae of WT, Gstm1−/−, and Gstt1−/− mice. Data are expressed as the mean ± SEM. WT, wild-type.

Article Snippet: The following primary antibodies were used: Gstm1 (rabbit polyclonal, 1:4000, Proteintech, 12412- 1- AP), Gstt1 (rabbit polyclonal, 1:2000, Proteintech, 15838- 1- AP). p- P38 (rabbit polyclonal, 1:1000, Cell Signaling Technology, 9211S), p- JNK (rabbit polyclonal, 1:1000, Cell Signaling Technology, 4668T), p- ERK (rabbit polyclonal, 1:2000, Cell Signaling Technology, 4370T), and pAkt (rabbit polyclonal, 1:1000, Cell Signaling Technology, 13038S).

Techniques: Knock-Out, Expressing

Journal: The FASEB Journal

Article Title: Gstm1/Gstt1 is essential for reducing cisplatin ototoxicity in CBA/CaJ mice

doi: 10.1096/fj.202200324r

Figure Lengend Snippet: FIGURE 4 More severe hearing loss in Gstm1/Gstt1-DKO mice than in WT mice after cisplatin treatment. (A) Body weights of WT and Gstm1/Gstt1-DKO mice during cisplatin treatment. n = 6. (B) ABR thresholds in cisplatin-treated WT and Gstm1/Gstt1-DKO mice. n = 5. (C) DPOAE thresholds in cisplatin-treated WT and Gstm1/Gstt1-DKO mice. n = 5. (D and E) Confocal images of cochlear hair cells and the statistics of OHC loss in cisplatin-treated WT and Gstm1/Gstt1-DKO mice. n = 3, Scale bar: 40 μm. *p < .05, ***p < .001. Data are expressed as the mean ± SEM. ABR, auditory brainstem response; DKO, double-knockout; DPOAE, distortion product otoacoustic emission; OHC, outer hair cell; WT, wild-type.

Article Snippet: The following primary antibodies were used: Gstm1 (rabbit polyclonal, 1:4000, Proteintech, 12412- 1- AP), Gstt1 (rabbit polyclonal, 1:2000, Proteintech, 15838- 1- AP). p- P38 (rabbit polyclonal, 1:1000, Cell Signaling Technology, 9211S), p- JNK (rabbit polyclonal, 1:1000, Cell Signaling Technology, 4668T), p- ERK (rabbit polyclonal, 1:2000, Cell Signaling Technology, 4370T), and pAkt (rabbit polyclonal, 1:1000, Cell Signaling Technology, 13038S).

Techniques: Double Knockout

Journal: The FASEB Journal

Article Title: Gstm1/Gstt1 is essential for reducing cisplatin ototoxicity in CBA/CaJ mice

doi: 10.1096/fj.202200324r

Figure Lengend Snippet: FIGURE 5 Assessment of SV thickness and SGN density after cisplatin treatment. (A) Morphology of SV in cisplatin-treated WT and Gstm1/Gstt1-DKO mice. Scale bar: 20 μm. (B) Morphology of SGNs in cisplatin-treated WT and Gstm1/Gstt1-DKO mice. Scale bar: 20 μm. (C) Assessment of SV thickness in cisplatin-treated WT and Gstm1/Gstt1-DKO mice. n = 5. (D) Assessment of SGN density in cisplatin- treated WT and Gstm1/Gstt1-DKO mice. n = 4. Data are expressed as the mean ± SEM. DKO, double-knockout; SGN, spiral ganglion neuron; SV, stria vascularis; WT, wild-type.

Article Snippet: The following primary antibodies were used: Gstm1 (rabbit polyclonal, 1:4000, Proteintech, 12412- 1- AP), Gstt1 (rabbit polyclonal, 1:2000, Proteintech, 15838- 1- AP). p- P38 (rabbit polyclonal, 1:1000, Cell Signaling Technology, 9211S), p- JNK (rabbit polyclonal, 1:1000, Cell Signaling Technology, 4668T), p- ERK (rabbit polyclonal, 1:2000, Cell Signaling Technology, 4370T), and pAkt (rabbit polyclonal, 1:1000, Cell Signaling Technology, 13038S).

Techniques: Double Knockout

Journal: The FASEB Journal

Article Title: Gstm1/Gstt1 is essential for reducing cisplatin ototoxicity in CBA/CaJ mice

doi: 10.1096/fj.202200324r

Figure Lengend Snippet: FIGURE 6 Assessment of GST activity and oxidative DNA and protein damage in the cochleae of cisplatin-treated mice. (A) Immunostaining of 4-HNE in the organ of Corti of cisplatin-treated WT and Gstm1/Gstt1-DKO mice (Green:4-HNE Blue: DAPI). Scale bar: 20 μm. (B) Relative intensity of 4-HNE to DAPI signal. n = 3. (C) Immunostaining of 3-NT in OHCs (Red: 3-NT Blue: DAPI). Scale bar: 20 μm. (D) Relative intensity of 3-NT to DAPI signal. n = 3. (E) Measurement of GST activity towards CDNB. n = 3. (F) The 8-OHdG expression level. n = 3. (G) Measurement of protein carbonyl in the cochleae of cisplatin-treated WT and Gstm1/Gstt1-DKO mice. n = 3. *p < .05, ***p < .001. Data are expressed as the mean ± SEM. 3-NT, 3-nitrotyrosine; 4-HNE, 4-hydroxynonenal; DKO, double-knockout; GST, glutathione S-transferase; OHC, outer hair cell; WT, wild-type.

Article Snippet: The following primary antibodies were used: Gstm1 (rabbit polyclonal, 1:4000, Proteintech, 12412- 1- AP), Gstt1 (rabbit polyclonal, 1:2000, Proteintech, 15838- 1- AP). p- P38 (rabbit polyclonal, 1:1000, Cell Signaling Technology, 9211S), p- JNK (rabbit polyclonal, 1:1000, Cell Signaling Technology, 4668T), p- ERK (rabbit polyclonal, 1:2000, Cell Signaling Technology, 4370T), and pAkt (rabbit polyclonal, 1:1000, Cell Signaling Technology, 13038S).

Techniques: Activity Assay, Immunostaining, Expressing, Double Knockout

Journal: The FASEB Journal

Article Title: Gstm1/Gstt1 is essential for reducing cisplatin ototoxicity in CBA/CaJ mice

doi: 10.1096/fj.202200324r

Figure Lengend Snippet: FIGURE 7 mRNA expression levels of detoxification genes in the cochleae of cisplatin-treated mice. (A) mRNA expression levels of genes involved in phase II detoxifying enzymatic activities in the cochleae of cisplatin-treated WT and Gstm1/Gstt1-DKO mice. n = 3, **p < .01, ***p < .001. (B) Nrf2 mRNA expression levels in the cochleae of cisplatin-treated WT and Gstm1/Gstt1-DKO mice. n = 3, ***p < .001. Data are expressed as the mean ± SEM; DKO, double-knockout; WT, wild-type.

Article Snippet: The following primary antibodies were used: Gstm1 (rabbit polyclonal, 1:4000, Proteintech, 12412- 1- AP), Gstt1 (rabbit polyclonal, 1:2000, Proteintech, 15838- 1- AP). p- P38 (rabbit polyclonal, 1:1000, Cell Signaling Technology, 9211S), p- JNK (rabbit polyclonal, 1:1000, Cell Signaling Technology, 4668T), p- ERK (rabbit polyclonal, 1:2000, Cell Signaling Technology, 4370T), and pAkt (rabbit polyclonal, 1:1000, Cell Signaling Technology, 13038S).

Techniques: Expressing, Double Knockout

Journal: The FASEB Journal

Article Title: Gstm1/Gstt1 is essential for reducing cisplatin ototoxicity in CBA/CaJ mice

doi: 10.1096/fj.202200324r

Figure Lengend Snippet: FIGURE 8 Increased apoptotic cell death in cisplatin-treated Gstm1/Gstt1-DKO mice. (A) Representative images of the TUNEL assay for OHCs in WT and Gstm1/Gstt1-DKO mice after cisplatin treatment. Scale bar: 40 μm. (B) Percentage of TUNEL-positive OHCs at specific regions of the organ of Corti. n = 5, *p < .05. (C and D) Analysis and quantifications of representative pro-survival protein. n = 3, *p < .05. Data are expressed as the mean ± SEM. DKO, double-knockout; Nrf2, nuclear transcription factor E2-related factor 2; OHC, outer hair cell; WT, wild-type.

Article Snippet: The following primary antibodies were used: Gstm1 (rabbit polyclonal, 1:4000, Proteintech, 12412- 1- AP), Gstt1 (rabbit polyclonal, 1:2000, Proteintech, 15838- 1- AP). p- P38 (rabbit polyclonal, 1:1000, Cell Signaling Technology, 9211S), p- JNK (rabbit polyclonal, 1:1000, Cell Signaling Technology, 4668T), p- ERK (rabbit polyclonal, 1:2000, Cell Signaling Technology, 4370T), and pAkt (rabbit polyclonal, 1:1000, Cell Signaling Technology, 13038S).

Techniques: TUNEL Assay, Double Knockout

Journal: Journal of Toxicologic Pathology

Article Title: Thioacetamide-induced Hepatocellular Necrosis Is Attenuated in Diet-induced Obese Mice

doi: 10.1293/tox.26.175

Figure Lengend Snippet: Western blots of hepatic drug-metabolizing enzymes in the livers of vehicle-treated mice. The number of mice fed the high-fat diet (HFD) or normal rodent diet (ND) was 4. A: Western blots of CYP2E1, glutathione S -transferases (GST) A1, GSTA2, GSTA3, GSTA4, GSTP1 and GSTM1 of HFD mice and ND mice are shown. B: Expression of drug-metabolizing enzymes as quantified by densitometry is shown. Black columns show the relative expression ratios of HFD mice to ND mice. *, ** p<0.05 or p<0.01 versus the expression in mice fed the normal rodent diet, respectively (Aspin-Welch t-test).

Article Snippet: To detect the drug-metabolizing enzyme expression, antibodies against CYP2E1 (Chemicon International, Temecula, CA, USA), GSTA1, GSTA2, GSTA3, GSTA4, GSTP1 and GSTM1 (Proteintech Group, Inc., Chicago, IL, USA) were used.

Techniques: Western Blot, Expressing

Journal: Basic and clinical andrology

Article Title: The protective effects of active ingredients from acrorus tatarinowii on sperm and their molecular mechanisms.

doi: 10.1186/s12610-024-00247-w

Figure Lengend Snippet: Fig. 3 Acrorus tatarinowii controls the BCL2-related pathway in the oligoasthenozoospermia rat. a-j qPCR analysis of mRNA expression of TNF-α, MAPK14, GSPT1, KDR, HMOX1, GSTM1, ESR1, CYP1B1, CYP1A1, and BCL2 in rat testicular tissues. K Western blot detection of protein expression of each gene. n = 3,vs.control,*P < 0.05,**P < 0.01,***P < 0.001 vs.Oligoasthenozoospermia,&P < 0.05,&&P < 0.01, &&&P < 0.001, ns, no significance

Article Snippet: Subsequently, proteins were transferred onto membranes and incubated with primary antibodies against TNF-α (CST), MAPK14 (CST), KDR (CST), HMOX1 (CST), GSTP1 (CST), GSTM1 (Proteintech), ESR1 (Proteintech), CYP1B1 (Proteintech), CYP1A1 (Proteintech), and BCL2 (Abcam).

Techniques: Expressing, Western Blot, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Wogonin alleviates liver injury in sepsis through Nrf2‐mediated NF‐κB signalling suppression

doi: 10.1111/jcmm.16604

Figure Lengend Snippet: Wogonin prevents hepatocytes from LPS/TNF‐α‐induced apoptosis. A, qRT‐PCR analysis of TNF‐α, IL‐1β and IL‐6 mRNA levels and (B) immunoblotting analysis of p65 and p‐p65 (Ser536) after LPS/TNF‐α and wogonin treatment with indicated concentrations in AML12 cells. C, Representative flow cytometry images and analysis of cell apoptosis after indicated treatment in AML12 cells. D, Immunoblotting analysis of pro‐apoptotic Bax, Cleaved‐Caspase3, Cleaved‐PARP and anti‐apoptotic Bcl2 expression related to (C). E, Densitometry analysis of Cleaved‐caspase3, Cleaved‐PARP, Bax and Bcl2 related to (D). NC, the negative control group. The results are presented as mean ± SD through at least 3 independent experiments and were analysed by one‐way analysis of variance (ANOVA). * P < .05, ** P < .01, *** P < .001

Article Snippet: The primary antibodies used for immunoblotting were against Nrf2 (rabbit polyclonal, ProteinTech, 16396‐1‐AP, 1:1000), p65 (rabbit polyclonal, ProteinTech, 10745‐1‐AP, 1:2000), p‐p65 (Ser536) (rabbit monoclonal, Cell Signaling Technology, #3033, 1:1000), β‐actin (mouse monoclonal, Cell Signaling Technology, #3700, 1:3000), NQO‐1 (rabbit polyclonal, Affinity Biosciences, DF6437, 1:500), GST‐M1 (rabbit polyclonal, ProteinTech, 12412‐1‐AP, 1:500), HO‐1 (rabbit polyclonal, Affinity Biosciences, AF5393, 1:500), SOD1 (rabbit polyclonal, ProteinTech, 10269‐1‐AP, 1:1000), SOD2 (mouse monoclonal, ProteinTech, 66474‐1‐Ig, 1:1000), Caspase3 (mouse monoclonal, ProteinTech, 66470‐2‐Ig, 1:1000), Cleaved‐Caspase3 (rabbit monoclonal, Cell Signaling Technology, #9664, 1:1000), Bax (rabbit polyclonal, ProteinTech, #50599‐2‐Ig, 1:2000), Bcl‐2 (rabbit polyclonal, ProteinTech, #12789‐1‐AP, 1:1000), PARP (rabbit monoclonal, Cell Signaling Technology, #9532, 1:1000), Cleaved‐PARP (Asp214) (rabbit monoclonal, Cell Signaling Technology, #5625, 1:500) and the second antibodies were HRP‐linked goat anti‐rabbit antibody (Cell Signaling Technology, #7074, 1:2000), HRP‐linked goat anti‐mouse antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Quantitative RT-PCR, Western Blot, Flow Cytometry, Expressing, Negative Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Wogonin alleviates liver injury in sepsis through Nrf2‐mediated NF‐κB signalling suppression

doi: 10.1111/jcmm.16604

Figure Lengend Snippet: Wogonin ameliorates septic liver injury in a Nrf2‐dependent manner. A and B, MDA and SOD activity in AML12 cells with indicated treatments. C, Intracellular ROS analysed by flow cytometry and represented as MFI of CM‐H 2 DCFDA in AML12 cells. D, Immunoblotting analysis of Nrf2, NQO‐1, GST‐M1, HO‐1, SOD1 and SOD2 in AML12 cells with indicated treatments. E and F, qRT‐PCR analysis of TNF‐α, IL‐1β and IL‐6 mRNA levels and immunoblotting analysis of p65 and p‐p65 (Ser536) in AML12 cells with indicated treatments. G, Representative flow cytometry images and analysis of cell apoptosis in AML12 cells with indicated treatments. H, Immunoblotting analysis of Nrf2, pro‐apoptotic Bax, Cleaved‐Caspase3 and Cleaved‐PARP together with anti‐apoptotic Bcl2 expression and densitometry analysis of Cleaved‐caspase3 and Cleaved‐PARP. The concentration of wogonin is 10 μmol/L. The results are presented as mean ± SD through at least 3 independent experiments and were analysed by two‐tailed Student's t test or one‐way analysis of variance (ANOVA). ns, no statistical significance, * P < .05, ** P < .01, *** P < .001

Article Snippet: The primary antibodies used for immunoblotting were against Nrf2 (rabbit polyclonal, ProteinTech, 16396‐1‐AP, 1:1000), p65 (rabbit polyclonal, ProteinTech, 10745‐1‐AP, 1:2000), p‐p65 (Ser536) (rabbit monoclonal, Cell Signaling Technology, #3033, 1:1000), β‐actin (mouse monoclonal, Cell Signaling Technology, #3700, 1:3000), NQO‐1 (rabbit polyclonal, Affinity Biosciences, DF6437, 1:500), GST‐M1 (rabbit polyclonal, ProteinTech, 12412‐1‐AP, 1:500), HO‐1 (rabbit polyclonal, Affinity Biosciences, AF5393, 1:500), SOD1 (rabbit polyclonal, ProteinTech, 10269‐1‐AP, 1:1000), SOD2 (mouse monoclonal, ProteinTech, 66474‐1‐Ig, 1:1000), Caspase3 (mouse monoclonal, ProteinTech, 66470‐2‐Ig, 1:1000), Cleaved‐Caspase3 (rabbit monoclonal, Cell Signaling Technology, #9664, 1:1000), Bax (rabbit polyclonal, ProteinTech, #50599‐2‐Ig, 1:2000), Bcl‐2 (rabbit polyclonal, ProteinTech, #12789‐1‐AP, 1:1000), PARP (rabbit monoclonal, Cell Signaling Technology, #9532, 1:1000), Cleaved‐PARP (Asp214) (rabbit monoclonal, Cell Signaling Technology, #5625, 1:500) and the second antibodies were HRP‐linked goat anti‐rabbit antibody (Cell Signaling Technology, #7074, 1:2000), HRP‐linked goat anti‐mouse antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Activity Assay, Flow Cytometry, Western Blot, Quantitative RT-PCR, Expressing, Concentration Assay, Two Tailed Test

Journal: Journal of Cellular and Molecular Medicine

Article Title: Wogonin alleviates liver injury in sepsis through Nrf2‐mediated NF‐κB signalling suppression

doi: 10.1111/jcmm.16604

Figure Lengend Snippet: Wogonin prevents hepatocytes from LPS/TNF‐α‐induced apoptosis. A, qRT‐PCR analysis of TNF‐α, IL‐1β and IL‐6 mRNA levels and (B) immunoblotting analysis of p65 and p‐p65 (Ser536) after LPS/TNF‐α and wogonin treatment with indicated concentrations in AML12 cells. C, Representative flow cytometry images and analysis of cell apoptosis after indicated treatment in AML12 cells. D, Immunoblotting analysis of pro‐apoptotic Bax, Cleaved‐Caspase3, Cleaved‐PARP and anti‐apoptotic Bcl2 expression related to (C). E, Densitometry analysis of Cleaved‐caspase3, Cleaved‐PARP, Bax and Bcl2 related to (D). NC, the negative control group. The results are presented as mean ± SD through at least 3 independent experiments and were analysed by one‐way analysis of variance (ANOVA). * P < .05, ** P < .01, *** P < .001

Article Snippet: The primary antibodies used for immunoblotting were against Nrf2 (rabbit polyclonal, ProteinTech, 16396‐1‐AP, 1:1000), p65 (rabbit polyclonal, ProteinTech, 10745‐1‐AP, 1:2000), p‐p65 (Ser536) (rabbit monoclonal, Cell Signaling Technology, #3033, 1:1000), β‐actin (mouse monoclonal, Cell Signaling Technology, #3700, 1:3000), NQO‐1 (rabbit polyclonal, Affinity Biosciences, DF6437, 1:500), GST‐M1 (rabbit polyclonal, ProteinTech, 12412‐1‐AP, 1:500), HO‐1 (rabbit polyclonal, Affinity Biosciences, AF5393, 1:500), SOD1 (rabbit polyclonal, ProteinTech, 10269‐1‐AP, 1:1000), SOD2 (mouse monoclonal, ProteinTech, 66474‐1‐Ig, 1:1000), Caspase3 (mouse monoclonal, ProteinTech, 66470‐2‐Ig, 1:1000), Cleaved‐Caspase3 (rabbit monoclonal, Cell Signaling Technology, #9664, 1:1000), Bax (rabbit polyclonal, ProteinTech, #50599‐2‐Ig, 1:2000), Bcl‐2 (rabbit polyclonal, ProteinTech, #12789‐1‐AP, 1:1000), PARP (rabbit monoclonal, Cell Signaling Technology, #9532, 1:1000), Cleaved‐PARP (Asp214) (rabbit monoclonal, Cell Signaling Technology, #5625, 1:500) and the second antibodies were HRP‐linked goat anti‐rabbit antibody (Cell Signaling Technology, #7074, 1:2000), HRP‐linked goat anti‐mouse antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Quantitative RT-PCR, Western Blot, Flow Cytometry, Expressing, Negative Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Wogonin alleviates liver injury in sepsis through Nrf2‐mediated NF‐κB signalling suppression

doi: 10.1111/jcmm.16604

Figure Lengend Snippet: Wogonin ameliorates septic liver injury in a Nrf2‐dependent manner. A and B, MDA and SOD activity in AML12 cells with indicated treatments. C, Intracellular ROS analysed by flow cytometry and represented as MFI of CM‐H 2 DCFDA in AML12 cells. D, Immunoblotting analysis of Nrf2, NQO‐1, GST‐M1, HO‐1, SOD1 and SOD2 in AML12 cells with indicated treatments. E and F, qRT‐PCR analysis of TNF‐α, IL‐1β and IL‐6 mRNA levels and immunoblotting analysis of p65 and p‐p65 (Ser536) in AML12 cells with indicated treatments. G, Representative flow cytometry images and analysis of cell apoptosis in AML12 cells with indicated treatments. H, Immunoblotting analysis of Nrf2, pro‐apoptotic Bax, Cleaved‐Caspase3 and Cleaved‐PARP together with anti‐apoptotic Bcl2 expression and densitometry analysis of Cleaved‐caspase3 and Cleaved‐PARP. The concentration of wogonin is 10 μmol/L. The results are presented as mean ± SD through at least 3 independent experiments and were analysed by two‐tailed Student's t test or one‐way analysis of variance (ANOVA). ns, no statistical significance, * P < .05, ** P < .01, *** P < .001

Article Snippet: The primary antibodies used for immunoblotting were against Nrf2 (rabbit polyclonal, ProteinTech, 16396‐1‐AP, 1:1000), p65 (rabbit polyclonal, ProteinTech, 10745‐1‐AP, 1:2000), p‐p65 (Ser536) (rabbit monoclonal, Cell Signaling Technology, #3033, 1:1000), β‐actin (mouse monoclonal, Cell Signaling Technology, #3700, 1:3000), NQO‐1 (rabbit polyclonal, Affinity Biosciences, DF6437, 1:500), GST‐M1 (rabbit polyclonal, ProteinTech, 12412‐1‐AP, 1:500), HO‐1 (rabbit polyclonal, Affinity Biosciences, AF5393, 1:500), SOD1 (rabbit polyclonal, ProteinTech, 10269‐1‐AP, 1:1000), SOD2 (mouse monoclonal, ProteinTech, 66474‐1‐Ig, 1:1000), Caspase3 (mouse monoclonal, ProteinTech, 66470‐2‐Ig, 1:1000), Cleaved‐Caspase3 (rabbit monoclonal, Cell Signaling Technology, #9664, 1:1000), Bax (rabbit polyclonal, ProteinTech, #50599‐2‐Ig, 1:2000), Bcl‐2 (rabbit polyclonal, ProteinTech, #12789‐1‐AP, 1:1000), PARP (rabbit monoclonal, Cell Signaling Technology, #9532, 1:1000), Cleaved‐PARP (Asp214) (rabbit monoclonal, Cell Signaling Technology, #5625, 1:500) and the second antibodies were HRP‐linked goat anti‐rabbit antibody (Cell Signaling Technology, #7074, 1:2000), HRP‐linked goat anti‐mouse antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Activity Assay, Flow Cytometry, Western Blot, Quantitative RT-PCR, Expressing, Concentration Assay, Two Tailed Test

Journal: Journal of Cellular and Molecular Medicine

Article Title: Wogonin alleviates liver injury in sepsis through Nrf2‐mediated NF‐κB signalling suppression

doi: 10.1111/jcmm.16604

Figure Lengend Snippet: Wogonin activates Nrf2 signalling in liver tissue of LPS‐induced sepsis mice. A‐E, Representative IHC images and IHC score analysis of Nrf2, HO‐1, SOD1, SOD2 and p‐p65 (Ser536) in liver tissue from LPS‐induced septic mice (n = 5). The number of scored fields = 10. Magnification, 400×. Scale bar = 50 μm. The concentration of wogonin was 50.0 mg/kg. The results are presented as mean ± SD through at least 3 independent experiments and were analysed by one‐way analysis of variance (ANOVA). ns, no statistical significance, * P < .05, ** P < .01, *** P < .001

Article Snippet: The primary antibodies used for immunoblotting were against Nrf2 (rabbit polyclonal, ProteinTech, 16396‐1‐AP, 1:1000), p65 (rabbit polyclonal, ProteinTech, 10745‐1‐AP, 1:2000), p‐p65 (Ser536) (rabbit monoclonal, Cell Signaling Technology, #3033, 1:1000), β‐actin (mouse monoclonal, Cell Signaling Technology, #3700, 1:3000), NQO‐1 (rabbit polyclonal, Affinity Biosciences, DF6437, 1:500), GST‐M1 (rabbit polyclonal, ProteinTech, 12412‐1‐AP, 1:500), HO‐1 (rabbit polyclonal, Affinity Biosciences, AF5393, 1:500), SOD1 (rabbit polyclonal, ProteinTech, 10269‐1‐AP, 1:1000), SOD2 (mouse monoclonal, ProteinTech, 66474‐1‐Ig, 1:1000), Caspase3 (mouse monoclonal, ProteinTech, 66470‐2‐Ig, 1:1000), Cleaved‐Caspase3 (rabbit monoclonal, Cell Signaling Technology, #9664, 1:1000), Bax (rabbit polyclonal, ProteinTech, #50599‐2‐Ig, 1:2000), Bcl‐2 (rabbit polyclonal, ProteinTech, #12789‐1‐AP, 1:1000), PARP (rabbit monoclonal, Cell Signaling Technology, #9532, 1:1000), Cleaved‐PARP (Asp214) (rabbit monoclonal, Cell Signaling Technology, #5625, 1:500) and the second antibodies were HRP‐linked goat anti‐rabbit antibody (Cell Signaling Technology, #7074, 1:2000), HRP‐linked goat anti‐mouse antibody (Cell Signaling Technology, #7076, 1:2000).

Techniques: Concentration Assay