Journal: Oxidative Medicine and Cellular Longevity

Article Title: Naringenin Alleviates Renal Ischemia Reperfusion Injury by Suppressing ER Stress-Induced Pyroptosis and Apoptosis through Activating Nrf2/HO-1 Signaling Pathway

doi: 10.1155/2022/5992436

Figure Lengend Snippet: Naringenin significantly inhibited pyroptosis and apoptosis induced by renal I/R injury in mice. (a) The levels of caspase-1 activity in the I/R group exhibited obvious reduction after naringenin treatment. (b–d) Naringenin significantly inhibited the mRNA expression of pyroptosis-related markers such as NLRP3, ASC, and caspase-1 in renal I/R injury. (e, f) Renal I/R-generated pyroptosis was obviously ameliorated by naringenin application as evidenced by the decreased protein expression of NLRP3, ASC, caspase-1, GSDMD-N, caspase-11, and IL-1 β in extracted kidney tissues. (g) The usage of naringenin remarkably restrained caspase-3 activity in renal I/R injury. (h) Western blot analysis utilized in protein detection of kidney tissue was selected to detect the protein levels of Bcl-2, BAX, and cleaved caspase-3 in the four groups. Values measured during animal experiments were carried out as mean ± SD, n = 3 (three times measurement). ∗ P < 0.05, compared with the Sham group; # P < 0.05, relative to the NS+I/R group.

Article Snippet: The dilutions and sources of all antibodies are as follows: GAPDH (10494-1-AP, 1 : 8000, Proteintech Group), GRP78 (11587-1-AP, 1 : 2000, Proteintech Group), CHOP (15204-1-AP, 1 : 2000, Proteintech Group), BAX (50599-2-Ig, 1 : 5000, Proteintech Group), cleaved caspase-3 (WL02117, 1 : 500, Wanleibio), HO-1 (10701-1-AP, 1 : 3000, Proteintech Group), Bcl-2 (26593-1-AP, 1 : 2000, Proteintech Group), Nrf2 (16396-1-AP, 1 : 5000, Proteintech Group), caspase-4 (sc-56056, 1 : 200, Santa Cruz), NLRP3 (#15101, 1 : 1000, Cell Signaling Technology), caspase-11 (sc-56038, 1 : 400, Santa Cruz), cleaved caspase-1 (sc-56036, 1 : 400, Santa Cruz), caspase-12 (sc-21747, 1 : 400, Santa Cruz), mature IL-1 β (#12242, 1 : 1000, Cell Signaling Technology), ASC (sc-514414, 1 : 200, Santa Cruz), GSDMD-N (#39754, 1 : 1000, Cell Signaling Technology), and KIM-1 (AF1817, MAB1750, 1 : 1000, R&D Systems).

Techniques: Activity Assay, Expressing, Generated, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Naringenin Alleviates Renal Ischemia Reperfusion Injury by Suppressing ER Stress-Induced Pyroptosis and Apoptosis through Activating Nrf2/HO-1 Signaling Pathway

doi: 10.1155/2022/5992436

Figure Lengend Snippet: Renal I/R-generated pyroptosis and apoptosis could be regulated by ER stress in mice. Mice in the 4-PBA+I/R group were injected intraperitoneally with 4-PBA (100 mg/kg, dilution in phosphate-buffered saline) 24 h before undergoing renal I/R surgery. (a, b) The levels of serum Cr and serum BUN in renal I/R injury decreased notably after inhibiting ER stress by 4-PBA application. (c, d) Quantitative analysis of tubular injury scores and representative images of H&E staining in different groups. (e) KIM-1 protein levels were assessed by western blot analysis. (f) The established inhibitor 4-PBA effectively restrained the protein expression of GRP78, CHOP, and caspase-12 in renal I/R injury. (g) The inhibition of ER stress by 4-PBA tremendously depressed the caspase-1 activity in mice with renal I/R surgery. (h) The mRNA levels of NLRP3, ASC, and caspase-1 in the I/R group declined after the application of 4-PBA. (i, j) 4-PBA as one specific inhibitor of ER stress remarkably depressed the activation of pyroptosis-related protein markers including NLRP3, ASC, caspase-1, GSDMD-N, IL-1 β , and caspase-11. (k) The protein levels of apoptotic markers consisting of BAX, Bcl-2, and cleaved caspase-3. Values measured during animal experiments were carried out as mean ± SD, n = 3 − 5. ∗ P < 0.05, compared with the Sham group; # P < 0.05, relative to the NS+I/R group.

Article Snippet: The dilutions and sources of all antibodies are as follows: GAPDH (10494-1-AP, 1 : 8000, Proteintech Group), GRP78 (11587-1-AP, 1 : 2000, Proteintech Group), CHOP (15204-1-AP, 1 : 2000, Proteintech Group), BAX (50599-2-Ig, 1 : 5000, Proteintech Group), cleaved caspase-3 (WL02117, 1 : 500, Wanleibio), HO-1 (10701-1-AP, 1 : 3000, Proteintech Group), Bcl-2 (26593-1-AP, 1 : 2000, Proteintech Group), Nrf2 (16396-1-AP, 1 : 5000, Proteintech Group), caspase-4 (sc-56056, 1 : 200, Santa Cruz), NLRP3 (#15101, 1 : 1000, Cell Signaling Technology), caspase-11 (sc-56038, 1 : 400, Santa Cruz), cleaved caspase-1 (sc-56036, 1 : 400, Santa Cruz), caspase-12 (sc-21747, 1 : 400, Santa Cruz), mature IL-1 β (#12242, 1 : 1000, Cell Signaling Technology), ASC (sc-514414, 1 : 200, Santa Cruz), GSDMD-N (#39754, 1 : 1000, Cell Signaling Technology), and KIM-1 (AF1817, MAB1750, 1 : 1000, R&D Systems).

Techniques: Generated, Injection, Staining, Western Blot, Expressing, Inhibition, Activity Assay, Activation Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Naringenin Alleviates Renal Ischemia Reperfusion Injury by Suppressing ER Stress-Induced Pyroptosis and Apoptosis through Activating Nrf2/HO-1 Signaling Pathway

doi: 10.1155/2022/5992436

Figure Lengend Snippet: Naringenin considerably mitigated H/R-induced pyroptosis and apoptosis in vitro. (a) The obvious elevation of caspase-1 activity in H/R exposure was remarkably depressed after naringenin application in renal HK-2 cells. (b) The pretreatment of naringenin evidently lessened the mRNA levels of typical pyroptosis-related markers including ASC, NLRP3, and caspase-1 during H/R injury. (c, d) The usage of naringenin markedly decreased the protein levels of representative pyroptosis-related markers such as NLRP3, caspase-1, ASC, GSDMD-N, IL-1 β , and caspase-4 after H/R exposure. (e, f) The flow cytometry revealed that naringenin administration tremendously mitigated H/R-generated apoptotic HK-2 cells. (g) The caspase-3 activity in various groups. (h) Western blot analysis utilized in protein detection of HK-2 cells was performed to quantify the protein levels of Bcl-2, cleaved caspase-3, and BAX in various group. Values measured during cellular experiments were carried out as mean ± SD, n = 3. ∗ P < 0.05, compared with the control group; # P < 0.05, versus the DMSO+H/R group.

Article Snippet: The dilutions and sources of all antibodies are as follows: GAPDH (10494-1-AP, 1 : 8000, Proteintech Group), GRP78 (11587-1-AP, 1 : 2000, Proteintech Group), CHOP (15204-1-AP, 1 : 2000, Proteintech Group), BAX (50599-2-Ig, 1 : 5000, Proteintech Group), cleaved caspase-3 (WL02117, 1 : 500, Wanleibio), HO-1 (10701-1-AP, 1 : 3000, Proteintech Group), Bcl-2 (26593-1-AP, 1 : 2000, Proteintech Group), Nrf2 (16396-1-AP, 1 : 5000, Proteintech Group), caspase-4 (sc-56056, 1 : 200, Santa Cruz), NLRP3 (#15101, 1 : 1000, Cell Signaling Technology), caspase-11 (sc-56038, 1 : 400, Santa Cruz), cleaved caspase-1 (sc-56036, 1 : 400, Santa Cruz), caspase-12 (sc-21747, 1 : 400, Santa Cruz), mature IL-1 β (#12242, 1 : 1000, Cell Signaling Technology), ASC (sc-514414, 1 : 200, Santa Cruz), GSDMD-N (#39754, 1 : 1000, Cell Signaling Technology), and KIM-1 (AF1817, MAB1750, 1 : 1000, R&D Systems).

Techniques: In Vitro, Activity Assay, Flow Cytometry, Generated, Western Blot

Journal: Journal of Immunology Research

Article Title: miR-181a-5p Inhibits Pyroptosis in Sepsis-Induced Acute Kidney Injury through Downregulation of NEK7

doi: 10.1155/2022/1825490

Figure Lengend Snippet: Pyroptosis of the LPS-induced HK-2 cells. (a) Cell viability of HK-2 and LPS-induced HK-2 cells. (b) Morphology of LPS-induced HK-2 cells under optical microscope (200x). (c) Immunofluorescence staining of N-GSDMD and (d) percentage of N-GSDMD-positive cells. (e) Immunofluorescence staining of ASC and (f) percentage of ASC-positive cells. ∗∗ P < 0.01.

Article Snippet: Then, the proteins were transferred to the PVDF membrane, followed by blocking for 1 h. Primary antibodies including anti-caspase-1 (ab62698, 1 : 1000, Abcam), anti-ASC protein (13833, 1 : 1000, Cell Signaling Technology), anti-GSDMD (39754, 1 : 1000, Cell Signaling Technology), anti-NLRP3 (13158, 1 : 1000, Cell Signaling Technology), and GAPDH (ab8245, 1 : 3000, Abcam) and the corresponding secondary antibodies were used.

Techniques: Microscopy, Immunofluorescence, Staining

Journal: Journal of Immunology Research

Article Title: miR-181a-5p Inhibits Pyroptosis in Sepsis-Induced Acute Kidney Injury through Downregulation of NEK7

doi: 10.1155/2022/1825490

Figure Lengend Snippet: Pyroptosis of HK-2 cells promotes inflammation. (a) The protein expression of NLRP3, NEK7, ASC, active caspase-1, and N-GSDMD in HK-2 and LPS-induced HK-2 cells by Western blot, and (b–f) quantification by ImageJ. (g) The expression of inflammatory factors including IL-18, IL-1 β , and THF- α by ELISA. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Then, the proteins were transferred to the PVDF membrane, followed by blocking for 1 h. Primary antibodies including anti-caspase-1 (ab62698, 1 : 1000, Abcam), anti-ASC protein (13833, 1 : 1000, Cell Signaling Technology), anti-GSDMD (39754, 1 : 1000, Cell Signaling Technology), anti-NLRP3 (13158, 1 : 1000, Cell Signaling Technology), and GAPDH (ab8245, 1 : 3000, Abcam) and the corresponding secondary antibodies were used.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Immunology Research

Article Title: miR-181a-5p Inhibits Pyroptosis in Sepsis-Induced Acute Kidney Injury through Downregulation of NEK7

doi: 10.1155/2022/1825490

Figure Lengend Snippet: Pyroptosis of renal tubular epithelial cells in the CLP-induced S-AKI mouse model. (a) The expression of caspase-1, N-GSDMD, NEK7, and NLRP3 in sham and CLP-induced mice by IHC. (b) The protein expression of NEK7, NLRP3, ASC, active caspase-1, and N-GSDMD by Western blot, and (c) quantification by ImageJ. (d) The expression of inflammatory factors including IL-18, IL-1 β , and THF- α . ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Then, the proteins were transferred to the PVDF membrane, followed by blocking for 1 h. Primary antibodies including anti-caspase-1 (ab62698, 1 : 1000, Abcam), anti-ASC protein (13833, 1 : 1000, Cell Signaling Technology), anti-GSDMD (39754, 1 : 1000, Cell Signaling Technology), anti-NLRP3 (13158, 1 : 1000, Cell Signaling Technology), and GAPDH (ab8245, 1 : 3000, Abcam) and the corresponding secondary antibodies were used.

Techniques: Expressing, Western Blot

Journal: Journal of Immunology Research

Article Title: miR-181a-5p Inhibits Pyroptosis in Sepsis-Induced Acute Kidney Injury through Downregulation of NEK7

doi: 10.1155/2022/1825490

Figure Lengend Snippet: miR-181a-5p inhibits pyroptosis of the LPS-induced HK-2 cells through downregulation of NEK7. (a) Morphology of LPS-induced HK-2 cells transfected with four vectors under an optical microscope (200x). The red arrow indicates pyroptosis of the cell. (b) Cell apoptosis of LPS-induced HK-2 cells transfected with four vectors by TUNEL, and (c) relative intensity of fluorescence. Scale bar = 100 μ m. (d) Cell viability of LPS-induced HK-2 cells transfected with four vectors by CCK-8 assay. (e) The protein expression of NEK7, NLRP3, N-GSDMD, and active caspase-1 by Western blot, and (f) quantification by ImageJ. (g) The expression of inflammatory factors including IL-1 β and IL-18. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Then, the proteins were transferred to the PVDF membrane, followed by blocking for 1 h. Primary antibodies including anti-caspase-1 (ab62698, 1 : 1000, Abcam), anti-ASC protein (13833, 1 : 1000, Cell Signaling Technology), anti-GSDMD (39754, 1 : 1000, Cell Signaling Technology), anti-NLRP3 (13158, 1 : 1000, Cell Signaling Technology), and GAPDH (ab8245, 1 : 3000, Abcam) and the corresponding secondary antibodies were used.

Techniques: Transfection, Microscopy, TUNEL Assay, Fluorescence, CCK-8 Assay, Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Nur77 Deficiency Exacerbates Macrophage NLRP3 Inflammasome-Mediated Inflammation and Accelerates Atherosclerosis

doi: 10.1155/2022/2017815

Figure Lengend Snippet: ox-LDL activates NLRP3 inflammasome and induces IL-1 β -mediated inflammation in PMs. PMs were treated with ox-LDL at a gradient concentration of 0, 25, 50, and 100 μ g/ml for 24 h and then were collected for analysis. (a) The LDH activity in supernatant was assessed by LDH Cytotoxicity Assay Kit. The expression levels of NLRP3 inflammasome-related protein NLRP3 (b), c-caspase-1 (c), c-IL-1 β (d), and GSMD-N (e) were determined by Western blotting. (f) The relative mRNA level of NLRP3 and IL-1 β ; ∗ P <0.05 compared with the control group. Relative protein levels of NLRP3 (g), c-caspase-1 (h), c-IL-1 β (i) and GSMD-N (j). n = 3. Values are shown as the mean ± SEM. ∗ P <0.05.

Article Snippet: The following primary antibodies were used: anti-NLRP3 (Cat# ab214185, Abcam), anti-caspase-1 (Cat# 3866, Cell Signaling Technology), anti-cleaved caspase-1 (Cat# 3866, Cell Signaling Technology), anti-IL-1 β (Cat# AB41610, Absci), anti-cleaved IL-1 β (Cat# AB41610, Absci), anti-GSMD (Cat# 39754, Cell Signaling Technology), anti-cleaved GSMD (Cat# 39754, Cell Signaling Technology), and anti-Nur77 (Cat# 3960, Cell Signaling Technology).

Techniques: Concentration Assay, Activity Assay, LDH Cytotoxicity Assay, Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Nur77 Deficiency Exacerbates Macrophage NLRP3 Inflammasome-Mediated Inflammation and Accelerates Atherosclerosis

doi: 10.1155/2022/2017815

Figure Lengend Snippet: Nur77 deficiency exacerbates NLRP3 inflammasome-mediated inflammation in PMs. PMs from WT or Nur77−/− mice were stimulated with or without ox-LDL (100 μ g/ml) for 24 h and then were collected for analysis. (a) The LDH activity in supernatant was assessed by LDH Cytotoxicity Assay Kit. The expression levels of NLRP3 inflammasome-related proteins NLRP3 (b), c-caspase-1 (c), c-IL-1 β (d), and GSMD-N (e) were determined by Western blotting. Relative protein levels of NLRP3 (f), c-caspase-1 (g), c-IL-1 β (h), and GSMD-N (i). (j) The mice NLRP3 promoter reporters, blank PGL3, and pcDNA or Nur77 plasmid were transiently transfected into Raw264.7 cells; after 24 h, the dual-luciferase activity was measured. (k) Raw264.7 cells were treated with or without ox-LDL (100 μ g/ml) for 24 h; DNA fragments from the Raw264.7 cells that contain regions of the NLRP3 promoter were immunoprecipitated with the anti-Nur77 antibody. The expression was identified by q-PCR. n = 3. Values are shown as the mean ± SEM. ∗ P < 0.05.

Article Snippet: The following primary antibodies were used: anti-NLRP3 (Cat# ab214185, Abcam), anti-caspase-1 (Cat# 3866, Cell Signaling Technology), anti-cleaved caspase-1 (Cat# 3866, Cell Signaling Technology), anti-IL-1 β (Cat# AB41610, Absci), anti-cleaved IL-1 β (Cat# AB41610, Absci), anti-GSMD (Cat# 39754, Cell Signaling Technology), anti-cleaved GSMD (Cat# 39754, Cell Signaling Technology), and anti-Nur77 (Cat# 3960, Cell Signaling Technology).

Techniques: Activity Assay, LDH Cytotoxicity Assay, Expressing, Western Blot, Plasmid Preparation, Transfection, Luciferase, Immunoprecipitation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Nur77 Deficiency Exacerbates Macrophage NLRP3 Inflammasome-Mediated Inflammation and Accelerates Atherosclerosis

doi: 10.1155/2022/2017815

Figure Lengend Snippet: ox-LDL activates NLRP3 inflammasome and induces IL-1 β -mediated inflammation in PMs. PMs were treated with ox-LDL at a gradient concentration of 0, 25, 50, and 100 μ g/ml for 24 h and then were collected for analysis. (a) The LDH activity in supernatant was assessed by LDH Cytotoxicity Assay Kit. The expression levels of NLRP3 inflammasome-related protein NLRP3 (b), c-caspase-1 (c), c-IL-1 β (d), and GSMD-N (e) were determined by Western blotting. (f) The relative mRNA level of NLRP3 and IL-1 β ; ∗ P <0.05 compared with the control group. Relative protein levels of NLRP3 (g), c-caspase-1 (h), c-IL-1 β (i) and GSMD-N (j). n = 3. Values are shown as the mean ± SEM. ∗ P <0.05.

Article Snippet: The following primary antibodies were used: anti-NLRP3 (Cat# ab214185, Abcam), anti-caspase-1 (Cat# 3866, Cell Signaling Technology), anti-cleaved caspase-1 (Cat# 3866, Cell Signaling Technology), anti-IL-1 β (Cat# AB41610, Absci), anti-cleaved IL-1 β (Cat# AB41610, Absci), anti-GSMD (Cat# 39754, Cell Signaling Technology), anti-cleaved GSMD (Cat# 39754, Cell Signaling Technology), and anti-Nur77 (Cat# 3960, Cell Signaling Technology).

Techniques: Concentration Assay, Activity Assay, LDH Cytotoxicity Assay, Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Nur77 Deficiency Exacerbates Macrophage NLRP3 Inflammasome-Mediated Inflammation and Accelerates Atherosclerosis

doi: 10.1155/2022/2017815

Figure Lengend Snippet: Nur77 deficiency exacerbates NLRP3 inflammasome-mediated inflammation in PMs. PMs from WT or Nur77−/− mice were stimulated with or without ox-LDL (100 μ g/ml) for 24 h and then were collected for analysis. (a) The LDH activity in supernatant was assessed by LDH Cytotoxicity Assay Kit. The expression levels of NLRP3 inflammasome-related proteins NLRP3 (b), c-caspase-1 (c), c-IL-1 β (d), and GSMD-N (e) were determined by Western blotting. Relative protein levels of NLRP3 (f), c-caspase-1 (g), c-IL-1 β (h), and GSMD-N (i). (j) The mice NLRP3 promoter reporters, blank PGL3, and pcDNA or Nur77 plasmid were transiently transfected into Raw264.7 cells; after 24 h, the dual-luciferase activity was measured. (k) Raw264.7 cells were treated with or without ox-LDL (100 μ g/ml) for 24 h; DNA fragments from the Raw264.7 cells that contain regions of the NLRP3 promoter were immunoprecipitated with the anti-Nur77 antibody. The expression was identified by q-PCR. n = 3. Values are shown as the mean ± SEM. ∗ P < 0.05.

Article Snippet: The following primary antibodies were used: anti-NLRP3 (Cat# ab214185, Abcam), anti-caspase-1 (Cat# 3866, Cell Signaling Technology), anti-cleaved caspase-1 (Cat# 3866, Cell Signaling Technology), anti-IL-1 β (Cat# AB41610, Absci), anti-cleaved IL-1 β (Cat# AB41610, Absci), anti-GSMD (Cat# 39754, Cell Signaling Technology), anti-cleaved GSMD (Cat# 39754, Cell Signaling Technology), and anti-Nur77 (Cat# 3960, Cell Signaling Technology).

Techniques: Activity Assay, LDH Cytotoxicity Assay, Expressing, Western Blot, Plasmid Preparation, Transfection, Luciferase, Immunoprecipitation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Astragaloside IV Alleviates Cerebral Ischemia-Reperfusion Injury through NLRP3 Inflammasome-Mediated Pyroptosis Inhibition via Activating Nrf2

doi: 10.1155/2021/9925561

Figure Lengend Snippet: Effect of AST IV on the NLRP3/Caspase-1/GSDMD pathway in MCAO/R rats. (a) Western blot assay showing the expression levels of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N. (b)–(f) Quantitative analysis of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N levels normalized to β -actin. Values are mean ± SD, n = 5. ∗∗ P < 0.01 vs. the sham group; # P < 0.05, ## P < 0.01 vs. the MCAO/R group.

Article Snippet: After electrophoresis, membrane transfer, and blocking, the membrane was incubated with primary antibodies (NLRP3 [1 : 1000, Cell Signaling Technology, batch number: 13158S], Caspase-1[1 : 1000, Cell Signaling Technology, batch number: 2225S], IL-1 β [1 : 1000, Abcam, No. 660091.1.g], GSDMD [1 : 1000, Cell Signaling Technology, batch number: 39754S], Nrf2 [1 : 1000, Affinity Biosciences, batch number: AF0639], lamin B1 [1 : 1000, Cell Signaling Technology, batch number: 17416S], and β -actin [1 : 8000, Sigma, batch number: A5441) at 4°C overnight.

Techniques: Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Astragaloside IV Alleviates Cerebral Ischemia-Reperfusion Injury through NLRP3 Inflammasome-Mediated Pyroptosis Inhibition via Activating Nrf2

doi: 10.1155/2021/9925561

Figure Lengend Snippet: Effect of AST IV on the NLRP3/Caspase-1/GSDMD pathway in OGD/R SH-SY5Y cells. (a) Western blot assay showing the expression levels of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N. (b)–(f) Quantitative analysis of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N levels normalized to β -actin. Values are mean ± SD, n = 3. ∗∗ P < 0.01 vs. the sham group; # P < 0.05, ## P < 0.01 vs. the OGD/R group.

Article Snippet: After electrophoresis, membrane transfer, and blocking, the membrane was incubated with primary antibodies (NLRP3 [1 : 1000, Cell Signaling Technology, batch number: 13158S], Caspase-1[1 : 1000, Cell Signaling Technology, batch number: 2225S], IL-1 β [1 : 1000, Abcam, No. 660091.1.g], GSDMD [1 : 1000, Cell Signaling Technology, batch number: 39754S], Nrf2 [1 : 1000, Affinity Biosciences, batch number: AF0639], lamin B1 [1 : 1000, Cell Signaling Technology, batch number: 17416S], and β -actin [1 : 8000, Sigma, batch number: A5441) at 4°C overnight.

Techniques: Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Astragaloside IV Alleviates Cerebral Ischemia-Reperfusion Injury through NLRP3 Inflammasome-Mediated Pyroptosis Inhibition via Activating Nrf2

doi: 10.1155/2021/9925561

Figure Lengend Snippet: Effect of AST IV on the NLRP3/Caspase-1/GSDMD pathway via Nrf2 in MCAO/R rats. (a) Western blot assay showing the expression levels of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N. (b)–(f) Quantitative analysis of the expression levels of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N normalized to β -actin. Values are mean ± SD, n = 5. ∗∗ P < 0.01 vs. the Sham group; ## P < 0.01 vs. the MCAO/R group; && P < 0.01 vs. the AST IV group.

Article Snippet: After electrophoresis, membrane transfer, and blocking, the membrane was incubated with primary antibodies (NLRP3 [1 : 1000, Cell Signaling Technology, batch number: 13158S], Caspase-1[1 : 1000, Cell Signaling Technology, batch number: 2225S], IL-1 β [1 : 1000, Abcam, No. 660091.1.g], GSDMD [1 : 1000, Cell Signaling Technology, batch number: 39754S], Nrf2 [1 : 1000, Affinity Biosciences, batch number: AF0639], lamin B1 [1 : 1000, Cell Signaling Technology, batch number: 17416S], and β -actin [1 : 8000, Sigma, batch number: A5441) at 4°C overnight.

Techniques: Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Astragaloside IV Alleviates Cerebral Ischemia-Reperfusion Injury through NLRP3 Inflammasome-Mediated Pyroptosis Inhibition via Activating Nrf2

doi: 10.1155/2021/9925561

Figure Lengend Snippet: Effect of AST IV on the NLRP3/Caspase-1/GSDMD pathway via Nrf2 in OGD/R SH-SY5Y cells. (a) Western blot assay showing the expression levels of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N. (b)–(f) Quantitative analysis of the expression levels of NLRP3, Caspase-1, IL-1 β , GSDMD, and GSDMD-N normalized to β -actin. Values are mean ± SD, n = 3. ∗ P < 0.05. ∗∗ P < 0.01 vs. the sham group; # P < 0.05, ## P < 0.01 vs. the OGD/R group; & P < 0.05, && P < 0.01 vs. the AST IV group.

Article Snippet: After electrophoresis, membrane transfer, and blocking, the membrane was incubated with primary antibodies (NLRP3 [1 : 1000, Cell Signaling Technology, batch number: 13158S], Caspase-1[1 : 1000, Cell Signaling Technology, batch number: 2225S], IL-1 β [1 : 1000, Abcam, No. 660091.1.g], GSDMD [1 : 1000, Cell Signaling Technology, batch number: 39754S], Nrf2 [1 : 1000, Affinity Biosciences, batch number: AF0639], lamin B1 [1 : 1000, Cell Signaling Technology, batch number: 17416S], and β -actin [1 : 8000, Sigma, batch number: A5441) at 4°C overnight.

Techniques: Western Blot, Expressing