Journal: Redox biology

Article Title: Ubiquitination as a key regulatory mechanism for O 3 -induced cutaneous redox inflammasome activation.

doi: 10.1016/j.redox.2022.102440

Figure Lengend Snippet: Fig. 1. O3 activated NLRP1 inflammasome via Caspase I (a) Immunofluorescence staining for NLRP1 (green) and ASC (red) in HaCaT cells silenced for NLRP1 10 nM for 24 h and exposed to O3. The blue staining (DAPI) represents nuclei. Images were taken at 40 × magnification (scale bar = 40 μm) and the fluorescent signal was quantified using ImageJ software. (b) Protein expression levels and relative quantification graph of p20 Caspase 1 over pro- Caspase 1 in HaCaT cells silenced for NLRP1 10 nM and then exposed to O3. (c) IL-1β released levels in media of HaCaT cells silenced for NLRP1 10 nM for 24 h and then exposed to O3. (d) Protein expression levels of p20 Caspase 1 over pro-Caspase 1 in HaCaT cells pre-treated with Caspase 1 inhibitor Z-YVAD- fmk 2 and 10 μM for 1 h and then exposed to O3. IL-1β mRNA expression levels (e) and released levels of IL- 1β (f) in HaCaT cells pre-treated with Caspase 1 in hibitor Z-YVAD-fmk 2 μM for 1 h and then exposed to O3. For all the experiments HaCaT cells were exposed to 0.4 ppm O3 for 1 h and then collected at the indicated timepoints post 1-h O3 exposure. For all western blotting the protein expression level was quantified using ImageJ software and β-actin was used as internal control. Data are the results of the averages of at least three different experiments, *p < 0.05 and #p < 0.05 by 2-way ANOVA followed by Tukey’s post-hoc comparison test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies ASC (Cat. NBP1-78977 NovusBio, USA) 1:1000, Caspase 1 (2225S Cell Signaling Technology, Danvers, MA, USA) 1:1000, NLRP1 (AB-84361, SIC, Rome, Italy) 1:1000, Ubiquitin (ab7780, Abcam, USA) 1:2000, DPP9 (ab42080, Abcam, USA) 1:1000, 4HNE (ab46545, Abcam, USA), UBR2 (18852-1-AP Proteintech, USA) in TBS-T with 1% non-fat milk (Bio-Rad Laboratories, Inc., USA).

Techniques: Immunofluorescence, Staining, Software, Expressing, Quantitative Proteomics, Western Blot, Control, Comparison

Journal: Redox biology

Article Title: Ubiquitination as a key regulatory mechanism for O 3 -induced cutaneous redox inflammasome activation.

doi: 10.1016/j.redox.2022.102440

Figure Lengend Snippet: Fig. 2. NLRP1 inflammasome activation by H2O2 (a) H2O2 levels production in media of HaCaT cells pre-treated with 1000 U/ml of Catalase (Cat) for 2 h and then exposed to O3 assessed by AmplexRed assay. (b) mRNA expression levels of NLRP1 (upper panel) and ASC (bottom panel) in HaCaT cells pre-treated with 1000 U/ml of Cat for 2 h and then exposed to O3. (c) Double Immunofluorescence staining for NLRP1 (red) and ASC (green) in HaCaT cells pre- treated with Cat 1000 U/ml for 2 h and then exposed to O3. Blue staining (DAPI) represents nuclei. Images were taken at 100 × magnification (scale bar = 2.5 μm); the fluorescent levels were quantified using ImageJ software. (d) Caspase 1 released levels in media of HaCaT cells pre-treated with Cat 1000 U/ ml for 2 h and then exposed to O3. (e) IL-1β mRNA expression levels in HaCaT cells pre-treated with Cat 1000 U/ml for 2 h and then exposed to O3. (f) Released levels of IL-1β in media of HaCaT cells pre- treated with Cat 1000 U/ml for 2 h and then exposed to O3. For all the experiments HaCaT cells were exposed to 0.4 ppm O3 for 1 h. Samples were collected at the indicated timepoints post 1-h O3 exposure. For all western blotting the protein expression level was quantified using ImageJ soft ware and β-actin was used as internal control. Data are the results of the averages of at least three different experiments, *p < 0.05 and #p < 0.05 by 2- way ANOVA followed by Tukey’s post-hoc compari son test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies ASC (Cat. NBP1-78977 NovusBio, USA) 1:1000, Caspase 1 (2225S Cell Signaling Technology, Danvers, MA, USA) 1:1000, NLRP1 (AB-84361, SIC, Rome, Italy) 1:1000, Ubiquitin (ab7780, Abcam, USA) 1:2000, DPP9 (ab42080, Abcam, USA) 1:1000, 4HNE (ab46545, Abcam, USA), UBR2 (18852-1-AP Proteintech, USA) in TBS-T with 1% non-fat milk (Bio-Rad Laboratories, Inc., USA).

Techniques: Activation Assay, Expressing, Double Immunofluorescence Staining, Staining, Software, Western Blot, Control

Journal: Redox biology

Article Title: Ubiquitination as a key regulatory mechanism for O 3 -induced cutaneous redox inflammasome activation.

doi: 10.1016/j.redox.2022.102440

Figure Lengend Snippet: Fig. 3. Involvement of 4HNE protein adducts in NLRP1 inflammasome activation (a) 4HNE protein expression levels in HaCaT cells exposed at the indi cated doses of O3 for 1 h and collected right after the end of O3 exposure. (b) Immunoprecipitation assay for NLRP1 probed with 4HNE in HaCaT cells pre- treated with proteasome inhibitor MG-132 20 μM for 2 h and exposed to O3 or H2O2 50 μM for 1 h. Samples were collected right after the end of 1-h O3/ H2O2 exposures. (c) Double Immunofluorescence staining for NLRP1 (red) and 4HNE (green) in HaCaT cells after O3 exposure. The blue staining (DAPI) represents nuclei. Images were taken at 60 × magnification (scale bar = 10 μm) and the fluorescent levels were quantified using ImageJ software. (d) Quantification of protein expression levels of ubiq uitinated proteins in HaCaT cells exposed to O3. (e) Protein expression levels of ubiquitinated-NLRP1 after Immunoprecipitation for NLRP1 in HaCaT cells pre-treated with proteasome inhibitor MG-132 20 μM for 2 h and Cat 1000 U/ml for 2 h and exposed to O3 right after the end of exposure. For all the experi ments HaCaT cells were exposed to 0.4 ppm O3 for 1 h and then collected at the indicated timepoints post 1-h O3 exposure. For all western blotting the protein expression level was quantified using ImageJ soft ware and β-actin was used as internal control. Data are the results of the averages of at least three different experiments, *p < 0.05 and #p < 0.05 by 2- way ANOVA followed by Tukey’s post-hoc compari son test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies ASC (Cat. NBP1-78977 NovusBio, USA) 1:1000, Caspase 1 (2225S Cell Signaling Technology, Danvers, MA, USA) 1:1000, NLRP1 (AB-84361, SIC, Rome, Italy) 1:1000, Ubiquitin (ab7780, Abcam, USA) 1:2000, DPP9 (ab42080, Abcam, USA) 1:1000, 4HNE (ab46545, Abcam, USA), UBR2 (18852-1-AP Proteintech, USA) in TBS-T with 1% non-fat milk (Bio-Rad Laboratories, Inc., USA).

Techniques: Activation Assay, Expressing, Immunoprecipitation, Double Immunofluorescence Staining, Staining, Software, Western Blot, Control

Journal: Redox biology

Article Title: Ubiquitination as a key regulatory mechanism for O 3 -induced cutaneous redox inflammasome activation.

doi: 10.1016/j.redox.2022.102440

Figure Lengend Snippet: Fig. 4. NLRP1 activation is mediated by its ubiq uitination. Protein expression levels of NLRP1 (a) in HaCaT cells pre-treated or not with the proteasome inhibitor MG-132 and then exposed to O3. (b) Double immunofluorescence staining for NLRP1 (green) and ASC (red), in HaCaT cells pre-treated with MG-132 and then exposed to O3. Blue staining (DAPI) repre sents nuclei. Images were taken at 40 × magnification (scale bar = 20 μm). The fluorescent levels were quantified using ImageJ software. Protein expression levels of p20 Caspase 1 over Pro-Caspase 1 (c), and UBR2 (d) in HaCaT cells pre-treated with MG-132 and exposed to O3. (e) Double IF staining for NLRP1 (red) and UBR2 (green) in HaCaT cells pre- treated with MG-132 and exposed to O3. Blue stain ing (DAPI) represents nuclei. Images were taken at 40 × magnification (scale bar = 20 μm). For all the experiments HaCaT cells were pre-treated or not with the proteasome inhibitor MG-132 20 μM for 2 h and then exposed to O3 0.4 ppm for 1 h. Samples were collected at the indicated timepoints post 1-h O3 exposure. For all western blotting the protein expression level was quantified using ImageJ soft ware and β-actin or Red ponceau were used as inter nal control. Data are the results of the averages of at least three different experiments, *p < 0.05 and #p < 0.05 by 2-way ANOVA followed by Tukey’s post-hoc comparison test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Membranes were incubated overnight at 4◦C with primary antibodies ASC (Cat. NBP1-78977 NovusBio, USA) 1:1000, Caspase 1 (2225S Cell Signaling Technology, Danvers, MA, USA) 1:1000, NLRP1 (AB-84361, SIC, Rome, Italy) 1:1000, Ubiquitin (ab7780, Abcam, USA) 1:2000, DPP9 (ab42080, Abcam, USA) 1:1000, 4HNE (ab46545, Abcam, USA), UBR2 (18852-1-AP Proteintech, USA) in TBS-T with 1% non-fat milk (Bio-Rad Laboratories, Inc., USA).

Techniques: Activation Assay, Expressing, Double Immunofluorescence Staining, Staining, Software, Western Blot, Control, Comparison

Journal: Biomolecules

Article Title: Albendazole-Schisandrin B Co-Therapy on Angiostrongylus cantonensis -Induced Meningoencephalitis in Mice

doi: 10.3390/biom10071001

Figure Lengend Snippet: Primer pairs of candidate genes used in real time quantitative PCR.

Article Snippet: Membranes were blocked with 5% nonfat milk prior to overnight incubation at 4 °C with agitation with the following primary antibodies: α-tubulin (GTX628802, GeneTex, Irvine, CA, USA), NLRP1B (SC-390133, Santa Cruz Biotechnology, Dallas, TX, USA), NLRC4 (06-1125, EMD Millipore), caspase-1 (22915-1-AP, Proteintech, Rosemont, IL, USA), IL-18 (10663-1-AP, Proteintech, Rosemont, IL, USA), IL-1β (16806-1-AP, Proteintech, Rosemont, IL, USA),Caspase-3 (GTX110543,GeneTex, Irvine, CA, USA), Caspase-8 (GTX110723, GeneTex, Irvine, CA, USA), Caspase-9 (GTX112888, GeneTex, Irvine, CA, USA), and BCL-2 (GTX100064, GeneTex, Irvine, CA, USA).

Techniques:

Journal: Biomolecules

Article Title: Albendazole-Schisandrin B Co-Therapy on Angiostrongylus cantonensis -Induced Meningoencephalitis in Mice

doi: 10.3390/biom10071001

Figure Lengend Snippet: Alb-Sch B co-therapy inhibits inflammasome activation and pyroptosis in A. cantonensis -infected mice. ( A ) RNA transcription levels of inflammasome components, including NLRC4, NLRP1B, Caspase-1, IL-18, IL-1β, and gasdermin D (GSDMD),were measured by qPCR. Data are presented as the mean ± SD (n = 5). ( B ) Representative western blot images reflecting protein levels ofinflammasome components. ( C ) Bar graphs showing protein expression levels relative to that of α-tubulin. Results are presented as the mean ± SD (n = 3). ( D ) Lactate dehydrogenase (LDH)levels measured in the cerebrospinal fluid (CSF), shown as the mean ± SD (n = 5). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001, and **** p -value < 0.0001.

Article Snippet: Membranes were blocked with 5% nonfat milk prior to overnight incubation at 4 °C with agitation with the following primary antibodies: α-tubulin (GTX628802, GeneTex, Irvine, CA, USA), NLRP1B (SC-390133, Santa Cruz Biotechnology, Dallas, TX, USA), NLRC4 (06-1125, EMD Millipore), caspase-1 (22915-1-AP, Proteintech, Rosemont, IL, USA), IL-18 (10663-1-AP, Proteintech, Rosemont, IL, USA), IL-1β (16806-1-AP, Proteintech, Rosemont, IL, USA),Caspase-3 (GTX110543,GeneTex, Irvine, CA, USA), Caspase-8 (GTX110723, GeneTex, Irvine, CA, USA), Caspase-9 (GTX112888, GeneTex, Irvine, CA, USA), and BCL-2 (GTX100064, GeneTex, Irvine, CA, USA).

Techniques: Activation Assay, Infection, Western Blot, Expressing

Journal: bioRxiv

Article Title: Oxidized thioredoxin-1 restrains the NLRP1 inflammasome

doi: 10.1101/2021.09.20.461118

Figure Lengend Snippet: ( A ) Schematic of the human NLRP1, CARD8, and NLRP3 proteins. The FIINDs of NLRP1 and CARD8 undergo autoproteolysis between the ZU5 and UPA subdomains. NT, N-terminus. CT, C-terminus. ( B ) HEK 293T cells were transfected with GFP- or NLRP1-FLAG. Lysates were subjected to anti-FLAG IP and quantitative MS analyses. The volcano plot depicts proteins enriched in the NLRP1-FLAG IP. ( C - E ) HEK 293T cells were transiently transfected with the indicated FLAG-tagged constructs, subjected to anti-FLAG IP, and analyzed by immunoblotting. In D , numbers indicate amino acid residues. An asterisk (*) denotes a background band.

Article Snippet: NLRP1 and CARD8 transcript variant 1 DNA obtained commercially from Origene (pCMV6-entry clones RC216481 and RC230245, respectively).

Techniques: Transfection, Construct, Western Blot

Journal: bioRxiv

Article Title: Oxidized thioredoxin-1 restrains the NLRP1 inflammasome

doi: 10.1101/2021.09.20.461118

Figure Lengend Snippet: ( A ) Cellular ROS levels were measured in CASP1 -/- N/TERT-1 keratinocytes (used to prevent pyroptosis) following treatment with the indicated (anti)oxidants using the cell permeable 2’,7’-dichlorodihydrofluorescein diacetate dye (DCFDA), which fluoresces upon oxidation. ( B ) Wild type N/TERT-1 keratinocytes treated with the indicated (anti)oxidants alone or together with VbP (10 µM) for 6 h before LDH and IL-1β release into supernatants was evaluated. ( D - F ) RAW 264.7 cells were treated with varying doses of the indicated (anti)oxidants alone or with VbP (10 µM) for 6 h before cell viability was assessed by Cell-TiterGlo (CTG, D ) and by LDH release assay ( E ) and immunoblot ( F ) for single dose (anti)oxidant concentration treatment. ( G ) MV4;11 wild type (top) and CARD8 -/- (bottom) cells were treated with varying doses of the indicated (anti)oxidants alone or with VbP (10 µM) for 6 h before cell viability was assessed by Cell-TiterGlo (CTG). Unless otherwise indicated, GSSG, GSH, DTT, NAC, TBHP, and H2O2 are treated at 100 µM, while Vit. K3, Antimycin A, VbP, and MitoTEMPO are treated at 10 µM, and Diamide was treated at 5 µM. All statistical data are means ± SEM for at least 3 biological replicates. * p < 0.05, * * p < 0.01, * * * p < 0.001.

Article Snippet: NLRP1 and CARD8 transcript variant 1 DNA obtained commercially from Origene (pCMV6-entry clones RC216481 and RC230245, respectively).

Techniques: Lactate Dehydrogenase Assay, Western Blot, Concentration Assay

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 expression is associated with senescence. A Human fibroblasts were exposed to 20 Gy ionizing irradiation (IR). On day 1, 5 and 7, NLRP1 and senescence protein expression were analyzed by immunoblotting. B Representative images of Ki67 and NLRP1 immunofluorescence. Scale bar = 50 μm. C , D IL-1β, IL18, IL-6 and IL-8 were quantified by ELISA. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, *** P < 0.001 differences between time points after irradiation and day 0. E , F Human fibroblasts were treated with Valboropro (Vbpro) to induce NLRP1 expression. After 24 h, NLRP1 and IL-6 protein expression were analyzed by immunoblotting and cytokines were analyzed by ELISA. Human fibroblasts were irradiated ( G ) or stimulated with palbociclib (Palbo) ( H ) to induce two different senescence models. Then, cells were transfected with a non-targeting control siRNA (Control) or with siRNAs against NLRP1 (siNLRP1). Expression of NLRP1 and senescence-associated proteins p16, p21, p53 and IL-6 were assessed by immunoblotting and IL6, IL-8 or IL-18 were quantified by ELISA. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, *** P < 0.001 irradiated vs. control. aaa P < 0.001, IR + siRNA vs. IR cells (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: Expressing, Irradiation, Western Blot, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Transfection, Control

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 is necessary for paracrine senescence. A Heat map depicting expression of 48 human cytokines in the medium at 7 days following culture from IR and IR + siRNA NLRP1. SASP was measured from the supernatant of the cells. B Effect of conditioned medium (CM) in human fibroblast growth. CM was collected from control, IR or IR + siNLRP1 cells. Percentage of cell growth was determined over 5 days. Image show cell populations stained with DAPI. C The induction of SA-β-Gal activity by CM was determined by microscopy. D IL-6 and IL-8 cytokine release to the medium after culture with CM from control, IR or IR + siNLRP1 cells. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, *** P < 0.001 irradiated vs. control. a P < 0.05, aa P < 0.005, aaa P < 0.001, IR + siRNA vs. IR cells (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: Expressing, Control, Staining, Activity Assay, Microscopy, Irradiation

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 contributes to cellular senescence in vivo. A Nlrp1 protein expression in liver from WT mice at 1 month after IR. B Serum levels of IL-18 after IR. C Effect of IR on the bodyweight of WT and Nlrp1 knockout (KO) mice. D Protein expression in liver from IR and non-IR WT and Nlrp1 KO mice of senescent markers (IL-6, p16 and p21). Densitometry in Supplementary Fig. 6. E Serum levels of IL-6 in serum from IR and non-IR WT and Nlrp1 KO mice. F Heat map depicting expression of 44 mouse cytokines in serum at 5 weeks after IR of WT and Nlrp1 KO mice. n = 6 mice per group. G IL-6 releases from healthy fibroblasts was assessed after 24 and 48 h of incubation with media containing serum from IR and non-IR WT and Nlrp1 KO mice. H Representative liver section stainings of hematoxylin and eosin (H&E). I Representative liver section immunostainings of NLRP1 and p16. All data are presented as means ± SEM, n = 6–8 mice per group; * P < 0.05, ** P < 0.005, *** P < 0.001 irradiated vs. control. aa P < 0.005 IR WT vs. IR KO mice; bb P < 0.005 IR KO vs. IR KO mice (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: In Vivo, Expressing, Knock-Out, Incubation, Irradiation, Control

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 is associated with senescence in humans. A β-Gal and Ki-67 immunostaining in liver section from a patient with a NLRP1 gain of function mutation. B Venn Diagrams showing genes that are changed in both selected datasets (left), and genes that are upregulated (middle) or downregulated (right). GSEA analysis showing two gene datasets enrichered for inflammatory response and targets of senescences genes is shown below (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: Immunostaining, Mutagenesis

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: NLRP1 senses DNA damage dependent of cGAS activation. A Protein expression levels of NLRP1 and cGAS after 24 h exposition to gDNA from non-irradiated and irradiated cells. B IL-6 and IL-18 release to the medium after the same experimental condition. Levels were determined by ELISA assay. C , D Protein expression levels of NLRP1 and cGAS and IL-6 and IL-18 release after 24 h exposition to a non-irradiated or irradiated synthetic double-stranded DNA sequence, poly(dA-dT), Cytokine levels were determined by ELISA assay. Cytokine levels were determined by ELISA assay. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, *** P < 0.001 gDNA vs. control cells. a P < 0.05, aa P < 0.005, aaa P < 0.001, IR gDNA vs. control cells. E Human fibroblasts were irradiated to induces senescence. Then, cells were transfected with a non-targeting control siRNA (Control) or with siRNAs against cGAS (sicGAS). Expression of NLRP1, NLRP3, IL-1β, cGAS and senescent protein p16, p21 was assessed by immunoblotting. F IL-18 release of non-irradiated, irradiated and irradiated and transfected with siRNAs against cGAS. Cytokine levels were determined by ELISA assay. All data are presented as means ± SEM, n = 4 independent experiments; ** P < 0.005, IR vs. control cells. aa P < 0.005, IR vs. sicGAS (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: Activation Assay, Expressing, Irradiation, Enzyme-linked Immunosorbent Assay, Sequencing, Control, Transfection, Western Blot

Journal: Inflammation Research

Article Title: NLRP1 inflammasome promotes senescence and senescence-associated secretory phenotype

doi: 10.1007/s00011-024-01892-7

Figure Lengend Snippet: Gasdermin D mediates SASP release. A Protein expression levels of GSDMD, its cleaved form, IL-6 and NLRP1 in fibroblasts which were irradiated (IR) and treated with necrosulfonamyde (NSA) for 5 days after IR. B , C IL-6 and IL-18 release determined by ELISA assay. Data are presented as means ± SEM, n = 5 independent experiments; *** P < 0.001 irradiated vs. control. aa P < 0.005 IR cells vs. IR NSA cells. D Heat map depicting expression of 44 mouse cytokines in serum at 5 weeks after IR of WT and Gsdmd KO mice n = 5 mice per group (color figure online)

Article Snippet: Monoclonal antibodies specific for NLRP1 (NBP1-54899), NLRP3 (NBP2-12446) and p21/CIP1/CDKN1A (NBP2-29463) were purchased from Novus Biologicals (Colorado, USA).

Techniques: Expressing, Irradiation, Enzyme-linked Immunosorbent Assay, Control