gsdmd n  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gsdmd n
    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, <t>GSDMD-N,</t> 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.
    Gsdmd N, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Naringenin Alleviates Renal Ischemia Reperfusion Injury by Suppressing ER Stress-Induced Pyroptosis and Apoptosis through Activating Nrf2/HO-1 Signaling Pathway"

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

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2022/5992436

    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.
    Figure Legend 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.

    Techniques Used: Activity Assay, Expressing, Generated, Western Blot

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

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

    gsdmd n  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gsdmd n
    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, <t>GSDMD-N,</t> 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.
    Gsdmd N, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    Images

    1) Product Images from "Naringenin Alleviates Renal Ischemia Reperfusion Injury by Suppressing ER Stress-Induced Pyroptosis and Apoptosis through Activating Nrf2/HO-1 Signaling Pathway"

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

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2022/5992436

    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.
    Figure Legend 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.

    Techniques Used: Activity Assay, Expressing, Generated, Western Blot

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

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

    anti gsdmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti gsdmd
    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 <t>N-GSDMD</t> and (d) percentage of N-GSDMD-positive cells. (e) Immunofluorescence staining <t>of</t> <t>ASC</t> and (f) percentage of ASC-positive cells. ∗∗ P < 0.01.
    Anti Gsdmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "miR-181a-5p Inhibits Pyroptosis in Sepsis-Induced Acute Kidney Injury through Downregulation of NEK7"

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

    Journal: Journal of Immunology Research

    doi: 10.1155/2022/1825490

    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.
    Figure Legend 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.

    Techniques Used: Microscopy, Immunofluorescence, Staining

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

    Techniques Used: Expressing, Western Blot

    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.
    Figure Legend 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.

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

    gasdermin d  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gasdermin d
    Gasdermin D, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti gsmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti gsmd
    ox-LDL activates NLRP3 inflammasome and induces <t>IL-1</t> <t>β</t> -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 <t>GSMD-N</t> (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.
    Anti Gsmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Nur77 Deficiency Exacerbates Macrophage NLRP3 Inflammasome-Mediated Inflammation and Accelerates Atherosclerosis"

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

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2022/2017815

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

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

    anti cleaved gsmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti cleaved gsmd
    ox-LDL activates NLRP3 inflammasome and induces <t>IL-1</t> <t>β</t> -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 <t>GSMD-N</t> (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.
    Anti Cleaved Gsmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Nur77 Deficiency Exacerbates Macrophage NLRP3 Inflammasome-Mediated Inflammation and Accelerates Atherosclerosis"

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

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2022/2017815

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

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

    gsdmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gsdmd
    Effect of AST IV on the <t>NLRP3/Caspase-1/GSDMD</t> pathway in MCAO/R rats. (a) Western blot assay showing the expression levels of NLRP3, Caspase-1, <t>IL-1</t> <t>β</t> , 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.
    Gsdmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Astragaloside IV Alleviates Cerebral Ischemia-Reperfusion Injury through NLRP3 Inflammasome-Mediated Pyroptosis Inhibition via Activating Nrf2"

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

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2021/9925561

    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.
    Figure Legend 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.

    Techniques Used: Western Blot, Expressing

    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.
    Figure Legend 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.

    Techniques Used: Western Blot, Expressing

    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.
    Figure Legend 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.

    Techniques Used: Western Blot, Expressing

    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.
    Figure Legend 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.

    Techniques Used: Western Blot, Expressing

    gasdermin d  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gasdermin d
    Gasdermin D, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    gasdermin d  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc gasdermin d
    Gasdermin D, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti gsdmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti gsdmd
    Baicalin prevents iohexol-induced pyroptosis of HK-2 cells by regulating inflammasome release. HK-2 cells were transfected with negative control siRNA (siNC) <t>or</t> <t>NLRP3</t> siRNA (siNLRP3), Caspase-1 siRNA (siCaspase-1). ( A ) The expression levels of NLRP3, ASC, Caspase-1 and <t>GSDMD</t> were detected by Western blotting. ( B ) Related quantification of NLRP3, ASC, Caspase-1 and GSDMD by grayscale analysis. a~c Means with the different letters are significantly different (p<005) by Duncan’s multiplerange tests.
    Anti Gsdmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Baicalin Alleviates Contrast-Induced Acute Kidney Injury Through ROS/NLRP3/Caspase-1/GSDMD Pathway-Mediated Proptosis in vitro"

    Article Title: Baicalin Alleviates Contrast-Induced Acute Kidney Injury Through ROS/NLRP3/Caspase-1/GSDMD Pathway-Mediated Proptosis in vitro

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S379629

    Baicalin prevents iohexol-induced pyroptosis of HK-2 cells by regulating inflammasome release. HK-2 cells were transfected with negative control siRNA (siNC) or NLRP3 siRNA (siNLRP3), Caspase-1 siRNA (siCaspase-1). ( A ) The expression levels of NLRP3, ASC, Caspase-1 and GSDMD were detected by Western blotting. ( B ) Related quantification of NLRP3, ASC, Caspase-1 and GSDMD by grayscale analysis. a~c Means with the different letters are significantly different (p<005) by Duncan’s multiplerange tests.
    Figure Legend Snippet: Baicalin prevents iohexol-induced pyroptosis of HK-2 cells by regulating inflammasome release. HK-2 cells were transfected with negative control siRNA (siNC) or NLRP3 siRNA (siNLRP3), Caspase-1 siRNA (siCaspase-1). ( A ) The expression levels of NLRP3, ASC, Caspase-1 and GSDMD were detected by Western blotting. ( B ) Related quantification of NLRP3, ASC, Caspase-1 and GSDMD by grayscale analysis. a~c Means with the different letters are significantly different (p<005) by Duncan’s multiplerange tests.

    Techniques Used: Transfection, Negative Control, Expressing, Western Blot

    anti gsdmd  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti gsdmd
    Change of the biomarkers and functional recovery in the SCI model after immunosuppressive therapy. (A) Statistical analysis of the BBB Scale in Sham, positive control, and treatment groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (B) Footprint analysis of different groups. (C, D) Statistical analysis of the Louisville Swim Scale in the four groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (E) Western blot showed that the expression level of the biomarkers in the treatment groups was between the levels of sham and control groups, and the levels of Bcl2 <t>and</t> <t>CASP3,</t> NFκB, <t>GSDMD,</t> IL1B, ASC, and CASP3 in SA and SV groups were also between those of Sham and control groups (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (F) RT-qPCR showed that the level of biomarkers was significantly decreased after treatment (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test).
    Anti Gsdmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Inhibition of IL1R1 or CASP4 attenuates spinal cord injury through ameliorating NLRP3 inflammasome-induced pyroptosis"

    Article Title: Inhibition of IL1R1 or CASP4 attenuates spinal cord injury through ameliorating NLRP3 inflammasome-induced pyroptosis

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2022.963582

    Change of the biomarkers and functional recovery in the SCI model after immunosuppressive therapy. (A) Statistical analysis of the BBB Scale in Sham, positive control, and treatment groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (B) Footprint analysis of different groups. (C, D) Statistical analysis of the Louisville Swim Scale in the four groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (E) Western blot showed that the expression level of the biomarkers in the treatment groups was between the levels of sham and control groups, and the levels of Bcl2 and CASP3, NFκB, GSDMD, IL1B, ASC, and CASP3 in SA and SV groups were also between those of Sham and control groups (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (F) RT-qPCR showed that the level of biomarkers was significantly decreased after treatment (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test).
    Figure Legend Snippet: Change of the biomarkers and functional recovery in the SCI model after immunosuppressive therapy. (A) Statistical analysis of the BBB Scale in Sham, positive control, and treatment groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (B) Footprint analysis of different groups. (C, D) Statistical analysis of the Louisville Swim Scale in the four groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (E) Western blot showed that the expression level of the biomarkers in the treatment groups was between the levels of sham and control groups, and the levels of Bcl2 and CASP3, NFκB, GSDMD, IL1B, ASC, and CASP3 in SA and SV groups were also between those of Sham and control groups (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (F) RT-qPCR showed that the level of biomarkers was significantly decreased after treatment (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test).

    Techniques Used: Functional Assay, Positive Control, Western Blot, Expressing, Quantitative RT-PCR

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    Cell Signaling Technology Inc gsdmd n
    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, <t>GSDMD-N,</t> 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.
    Gsdmd N, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti gsdmd
    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 <t>N-GSDMD</t> and (d) percentage of N-GSDMD-positive cells. (e) Immunofluorescence staining <t>of</t> <t>ASC</t> and (f) percentage of ASC-positive cells. ∗∗ P < 0.01.
    Anti Gsdmd, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc gasdermin d
    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 <t>N-GSDMD</t> and (d) percentage of N-GSDMD-positive cells. (e) Immunofluorescence staining <t>of</t> <t>ASC</t> and (f) percentage of ASC-positive cells. ∗∗ P < 0.01.
    Gasdermin D, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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