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R&D Systems human caspase
Human Caspase, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+caspase+1/Human+Caspase-1%2FICE+Quantikine+ELISA+Kit/pmc12865640-266-27-33
Average 94 stars, based on 63 article reviews

human caspase - by Bioz Stars, 2026-09

94/100 stars

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A01 suppressed NLRP3 inflammasome activation in BMDMs. (A–I) Bone marrow–derived macrophages (BMDMs) (100 ng/mL, 4 h) were primed with LPS, followed by incubation with A01 and stimulation with nigericin (10 µM, 30 min) (A–C), ATP (5 mM, 1 h) (D–F), or alum (300 μg/mL, 3 h) (G–I). Cleaved caspase-1 (p20) in the supernatant (Sup) and pro–caspase-1 in cell lysates (Lys) were examined by immunoblotting (A, D, G). IL-1β secretion in the supernatant was quantified by ELISA (B, E, H), and LDH release was measured to assess cell death (C, F, I). Numbers above the blots indicate densitometric ratios of cleaved caspase-1 (p20) to pro–caspase-1. Data represent mean ± SEM of triplicate samples and are representative of at least three independent experiments. ***p<0.001, compared with the LPS + nigericin/ATP/alum group (unpaired Student’s t -test).

Journal: Open Medicine

Article Title: Specific inhibition of NLRP3 inflammasome by a Smurf1 inhibitor in vitro and in vivo

doi: 10.1515/med-2026-1397

Figure Lengend Snippet: A01 suppressed NLRP3 inflammasome activation in BMDMs. (A–I) Bone marrow–derived macrophages (BMDMs) (100 ng/mL, 4 h) were primed with LPS, followed by incubation with A01 and stimulation with nigericin (10 µM, 30 min) (A–C), ATP (5 mM, 1 h) (D–F), or alum (300 μg/mL, 3 h) (G–I). Cleaved caspase-1 (p20) in the supernatant (Sup) and pro–caspase-1 in cell lysates (Lys) were examined by immunoblotting (A, D, G). IL-1β secretion in the supernatant was quantified by ELISA (B, E, H), and LDH release was measured to assess cell death (C, F, I). Numbers above the blots indicate densitometric ratios of cleaved caspase-1 (p20) to pro–caspase-1. Data represent mean ± SEM of triplicate samples and are representative of at least three independent experiments. ***p<0.001, compared with the LPS + nigericin/ATP/alum group (unpaired Student’s t -test).

Article Snippet: Anti-human caspase-1 (1:2000, 4199S) was purchased from Cell Signaling Technology.

Techniques: Activation Assay, Derivative Assay, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay

A01 had no impact on NLRC4 or AIM2 inflammasome activation. (A–F) LPS-primed BMDMs (100 ng/mL, 4 h) were treated with or without A01 and then transfected with poly (dA:dT) (5 μg/mL, 6 h) to activate AIM2 (A–C) or flagellin (1 μg/mL, 6 h) to activate NLRC4 (D–F). Cleaved caspase-1 (p20) in the supernatant and pro–caspase-1 in cell lysates were detected by immunoblotting (A, D). IL-1β secretion was measured by ELISA (B, E), and LDH release was assessed (C, F). Numbers above the blots indicate densitometric ratios of cleaved caspase-1 to pro–caspase-1. Data are presented as mean ± SEM of triplicates and represent at least three independent experiments.ns, not significant, compared with the LPS + poly (dA:dT) or LPS + flagellin group (unpaired Student’s t -test).

Journal: Open Medicine

Article Title: Specific inhibition of NLRP3 inflammasome by a Smurf1 inhibitor in vitro and in vivo

doi: 10.1515/med-2026-1397

Figure Lengend Snippet: A01 had no impact on NLRC4 or AIM2 inflammasome activation. (A–F) LPS-primed BMDMs (100 ng/mL, 4 h) were treated with or without A01 and then transfected with poly (dA:dT) (5 μg/mL, 6 h) to activate AIM2 (A–C) or flagellin (1 μg/mL, 6 h) to activate NLRC4 (D–F). Cleaved caspase-1 (p20) in the supernatant and pro–caspase-1 in cell lysates were detected by immunoblotting (A, D). IL-1β secretion was measured by ELISA (B, E), and LDH release was assessed (C, F). Numbers above the blots indicate densitometric ratios of cleaved caspase-1 to pro–caspase-1. Data are presented as mean ± SEM of triplicates and represent at least three independent experiments.ns, not significant, compared with the LPS + poly (dA:dT) or LPS + flagellin group (unpaired Student’s t -test).

Article Snippet: Anti-human caspase-1 (1:2000, 4199S) was purchased from Cell Signaling Technology.

Techniques: Activation Assay, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay

a Transcriptome profiling reveals elevated PYCARD expression in tumor (TNM stages I/II, n = 21; III, n = 33; IV, n = 29; combined I-IV, n = 83) versus non-tumor (N, n = 14) pancreatic tissues (VPCB cohort). CPM, counts per million. **** P < 0.0001; One-way ANOVA. b Kaplan-Meier survival of PDAC patients (VPCB, n = 82) stratified equally into low or high PYCARD expression. P -value indicated; Log-rank (Mantel-Cox) test. c Immunoblots for ASC, cleaved (p20) and pro (p45) Caspase-1, and Tubulin on representative PDAC patient tumor (T; stage III) and non-tumor pancreatic (N) tissue lysates. The samples were derived from the same experiment, but different gels for ASC and Tubulin, and another for Caspase-1 and Tubulin, were processed in parallel. Each lane represents an individual patient sample. Size of molecular weight markers (kD) is indicated. Blots represent at least 3 independent experiments. d Representative photomicrographs of human pancreatic non-tumor and PDAC tumor tissues immunostained for ASC. Scale bars = 60 μm. The graph depicts quantification of ASC-positive epithelial staining. n = 5/group. **** P < 0.0001; Student’s t-test, two-sided. Data are mean values ± SEM. e, f Immunofluorescence images (20× magnification) of ASC (red), cleaved (p10) Caspase-1 (violet), and CK19 (CK; panel e , green) or CD45 (panel f , green) staining on a PDAC patient tumor cross-section (representing at least 3 patient tumors). DAPI nuclear staining is blue. Also shown are 20× and 63× magnification merged images, the latter depicting regions within dotted white squares (20× images) showing nuclear localization of ASC (white staining), predominantly in cytokeratin-positive epithelium (panel e ). Arrows point to punctate intra- and extracellular ASC specks. Scale bars = 100 μm (20×) and 10μm (63×). g Representative photomicrographs of human pancreatic non-tumor and PDAC tumor tissues immunostained for cleaved (p10) Caspase-1. Scale bars = 60 μm. The graph depicts quantification of cleaved Caspase-1-positive epithelial staining. n = 5/group. ** P = 0.0013; Student’s t-test, two-sided. Data are mean values ± SEM. ELISAs for h total IL-18 in serum and i mature/free (p18) IL-18 in pancreatic tissue lysates from PDAC patients (VPCB). n = 10 ( h ) and 5 ( i ) per TNM stage group and cancer-free individuals (N). * P = 0.049 (N v III, h ), 0.0181 (N v I-IV, i ); ** P = 0.0019 (N v IV, h ), 0.0027 (N v I-IV, h ), 0.0014 (N v IV, i ); One-way ANOVA. Data are mean values ± SEM. j Transcriptome profiling of elevated IL18 gene expression in PDAC patient tumors (VPCB). *** P = 0.0006 (N v I/II), 0.0002 (N v III); **** P < 0.0001; One-way ANOVA. k Kaplan-Meier survival of PDAC patients (VPCB, n = 82) stratified equally into low or high IL18 gene expression. P -value indicated; Log-rank (Mantel-Cox) test. l Linear regression of tumoral PYCARD (ASC) or CASP1 mRNA and serum IL-18 protein levels in PDAC patients. r = Pearson’s correlation coefficient. P values indicated, two-sided test.

Journal: Nature Communications

Article Title: Cancer cell-intrinsic inflammasome protein ASC links innate immunity with mitochondrial metabolism in driving pancreatic cancer

doi: 10.1038/s41467-026-69398-w

Figure Lengend Snippet: a Transcriptome profiling reveals elevated PYCARD expression in tumor (TNM stages I/II, n = 21; III, n = 33; IV, n = 29; combined I-IV, n = 83) versus non-tumor (N, n = 14) pancreatic tissues (VPCB cohort). CPM, counts per million. **** P < 0.0001; One-way ANOVA. b Kaplan-Meier survival of PDAC patients (VPCB, n = 82) stratified equally into low or high PYCARD expression. P -value indicated; Log-rank (Mantel-Cox) test. c Immunoblots for ASC, cleaved (p20) and pro (p45) Caspase-1, and Tubulin on representative PDAC patient tumor (T; stage III) and non-tumor pancreatic (N) tissue lysates. The samples were derived from the same experiment, but different gels for ASC and Tubulin, and another for Caspase-1 and Tubulin, were processed in parallel. Each lane represents an individual patient sample. Size of molecular weight markers (kD) is indicated. Blots represent at least 3 independent experiments. d Representative photomicrographs of human pancreatic non-tumor and PDAC tumor tissues immunostained for ASC. Scale bars = 60 μm. The graph depicts quantification of ASC-positive epithelial staining. n = 5/group. **** P < 0.0001; Student’s t-test, two-sided. Data are mean values ± SEM. e, f Immunofluorescence images (20× magnification) of ASC (red), cleaved (p10) Caspase-1 (violet), and CK19 (CK; panel e , green) or CD45 (panel f , green) staining on a PDAC patient tumor cross-section (representing at least 3 patient tumors). DAPI nuclear staining is blue. Also shown are 20× and 63× magnification merged images, the latter depicting regions within dotted white squares (20× images) showing nuclear localization of ASC (white staining), predominantly in cytokeratin-positive epithelium (panel e ). Arrows point to punctate intra- and extracellular ASC specks. Scale bars = 100 μm (20×) and 10μm (63×). g Representative photomicrographs of human pancreatic non-tumor and PDAC tumor tissues immunostained for cleaved (p10) Caspase-1. Scale bars = 60 μm. The graph depicts quantification of cleaved Caspase-1-positive epithelial staining. n = 5/group. ** P = 0.0013; Student’s t-test, two-sided. Data are mean values ± SEM. ELISAs for h total IL-18 in serum and i mature/free (p18) IL-18 in pancreatic tissue lysates from PDAC patients (VPCB). n = 10 ( h ) and 5 ( i ) per TNM stage group and cancer-free individuals (N). * P = 0.049 (N v III, h ), 0.0181 (N v I-IV, i ); ** P = 0.0019 (N v IV, h ), 0.0027 (N v I-IV, h ), 0.0014 (N v IV, i ); One-way ANOVA. Data are mean values ± SEM. j Transcriptome profiling of elevated IL18 gene expression in PDAC patient tumors (VPCB). *** P = 0.0006 (N v I/II), 0.0002 (N v III); **** P < 0.0001; One-way ANOVA. k Kaplan-Meier survival of PDAC patients (VPCB, n = 82) stratified equally into low or high IL18 gene expression. P -value indicated; Log-rank (Mantel-Cox) test. l Linear regression of tumoral PYCARD (ASC) or CASP1 mRNA and serum IL-18 protein levels in PDAC patients. r = Pearson’s correlation coefficient. P values indicated, two-sided test.

Article Snippet: Immunoblotting was performed with antibodies against ASC (AdipoGen, AL177, 1:200), human Caspase-1 (Cell Signaling Technologies, D7F10 or 2225, 1:1000), mouse Caspase-1 (AdipoGen, AG-20B-0042-C100, 1:500), GSDMD (Abcam, ab219800, 1:2000) and α-Tubulin (Abcam, ab6160, 1:6667), as well as the OxPhos Rodent WB Antibody Cocktail (Invitrogen, 45-8099, 1:1000).

Techniques: Expressing, Western Blot, Derivative Assay, Molecular Weight, Staining, Immunofluorescence, Gene Expression

qPCR expression (normalized for 18 s rRNA ) of ( a ) Pycard (ASC) and Casp1 genes, and ( b ) inflammasome-associated pattern recognition receptor genes, in pancreas tissues of 20-week-old (wo) wild-type (WT) and KPC mice ( n = 6/genotype). * P = 0.035 ( Casp1 ), 0.0343 ( Mefv ), 0.0103 ( Nlrp3 ), 0.0255 ( Nlrp6 ); ** P = 0.0048 ( Pycard ), 0.0069 ( Aim2 ), 0.0040 ( Nlrc4 ); Student’s t-test, two-sided. Data are mean values ± SEM. c ASC, cleaved (p20) and pro (p45) Caspase-1, and Tubulin, immunoblots on 20wo WT and KPC mouse pancreas tissue lysates. The samples were derived from the same experiment but different gels for ASC and Tubulin, and another for Caspase-1, were processed in parallel. Each lane represents an individual mouse sample. The size of molecular weight markers (kD) is indicated. d Representative photomicrographs of 20wo WT and KPC pancreas tissue cross-sections immunostained for cleaved (p10) Caspase-1. Scale bars = 100 μm. Magnified image (inset in the KPC section) depicts nuclear and cytoplasmic staining in the ductal epithelium of a lesion. The graph shows quantification of cleaved Caspase-1-positive epithelial staining. n = 6/group. *** P = 0.0004; Student’s t-test, two-sided. Data are mean values ± SEM. e Images of DAPI (nuclear; blue), ASC (green), or cleaved (p10) Caspase-1 (red) immunofluorescence on 20wo WT (top row) and KPC (middle row) mouse pancreas cross-sections (representing 5 mice per genotype). Bottom row shows the corresponding merged staining image of 20wo WT and KPC mouse pancreas cross-sections. Yellow and white arrows point to representative cytosolic and nuclear, respectively, colocalized ASC “specks” with cleaved Caspase-1. Scale bars = 10 μm. f ELISAs for total IL-18 and IL-1β protein levels in serum from 20wo mice. n = 6/genotype. ** P = 0.0019; Student’s t-test, two-sided. Data are mean values ± SEM. g qPCR (normalized for 18 s rRNA ) of Il18 and Il1b genes in pancreas tissues of 20wo mice. n = 6/genotype. * P = 0.0315, ** P = 0.008; Student’s t test, two-sided. Data are mean values ± SEM. h Immunoblots of pancreas tissue lysates from 20wo mice for pro-IL-18 (p24) and pro-IL-1β (p31) (arrows), and Tubulin. Blots represent at least 3 independent experiments. The samples were derived from the same experiment, but different gels for IL-18 and Tubulin, and another for IL-1β and Tubulin, were processed in parallel. Each lane represents an individual mouse sample. The size of molecular weight markers (kD) is indicated. i ELISA for mature (p18) IL-18 protein levels in pancreas tissue lysates from 10wo and 20wo mice. n = 5/genotype. * P = 0.0116 (10wo), 0.0306 (20wo); Student’s t-test, two-sided. Data are mean values ± SEM.

Journal: Nature Communications

Article Title: Cancer cell-intrinsic inflammasome protein ASC links innate immunity with mitochondrial metabolism in driving pancreatic cancer

doi: 10.1038/s41467-026-69398-w

Figure Lengend Snippet: qPCR expression (normalized for 18 s rRNA ) of ( a ) Pycard (ASC) and Casp1 genes, and ( b ) inflammasome-associated pattern recognition receptor genes, in pancreas tissues of 20-week-old (wo) wild-type (WT) and KPC mice ( n = 6/genotype). * P = 0.035 ( Casp1 ), 0.0343 ( Mefv ), 0.0103 ( Nlrp3 ), 0.0255 ( Nlrp6 ); ** P = 0.0048 ( Pycard ), 0.0069 ( Aim2 ), 0.0040 ( Nlrc4 ); Student’s t-test, two-sided. Data are mean values ± SEM. c ASC, cleaved (p20) and pro (p45) Caspase-1, and Tubulin, immunoblots on 20wo WT and KPC mouse pancreas tissue lysates. The samples were derived from the same experiment but different gels for ASC and Tubulin, and another for Caspase-1, were processed in parallel. Each lane represents an individual mouse sample. The size of molecular weight markers (kD) is indicated. d Representative photomicrographs of 20wo WT and KPC pancreas tissue cross-sections immunostained for cleaved (p10) Caspase-1. Scale bars = 100 μm. Magnified image (inset in the KPC section) depicts nuclear and cytoplasmic staining in the ductal epithelium of a lesion. The graph shows quantification of cleaved Caspase-1-positive epithelial staining. n = 6/group. *** P = 0.0004; Student’s t-test, two-sided. Data are mean values ± SEM. e Images of DAPI (nuclear; blue), ASC (green), or cleaved (p10) Caspase-1 (red) immunofluorescence on 20wo WT (top row) and KPC (middle row) mouse pancreas cross-sections (representing 5 mice per genotype). Bottom row shows the corresponding merged staining image of 20wo WT and KPC mouse pancreas cross-sections. Yellow and white arrows point to representative cytosolic and nuclear, respectively, colocalized ASC “specks” with cleaved Caspase-1. Scale bars = 10 μm. f ELISAs for total IL-18 and IL-1β protein levels in serum from 20wo mice. n = 6/genotype. ** P = 0.0019; Student’s t-test, two-sided. Data are mean values ± SEM. g qPCR (normalized for 18 s rRNA ) of Il18 and Il1b genes in pancreas tissues of 20wo mice. n = 6/genotype. * P = 0.0315, ** P = 0.008; Student’s t test, two-sided. Data are mean values ± SEM. h Immunoblots of pancreas tissue lysates from 20wo mice for pro-IL-18 (p24) and pro-IL-1β (p31) (arrows), and Tubulin. Blots represent at least 3 independent experiments. The samples were derived from the same experiment, but different gels for IL-18 and Tubulin, and another for IL-1β and Tubulin, were processed in parallel. Each lane represents an individual mouse sample. The size of molecular weight markers (kD) is indicated. i ELISA for mature (p18) IL-18 protein levels in pancreas tissue lysates from 10wo and 20wo mice. n = 5/genotype. * P = 0.0116 (10wo), 0.0306 (20wo); Student’s t-test, two-sided. Data are mean values ± SEM.

Techniques: Expressing, Western Blot, Derivative Assay, Molecular Weight, Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay

a Representative 20-week-old (wo) wild-type (WT), KPC and KPC:ASC KO (KPC:KO) mouse pancreas images. Scale bars = 1 cm. b Total mass (grams; g) of pancreas in mice. n = 20 mice/group. **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. c Percentage area of whole pancreas (H&E-stained whole slide scans) containing lesions. n = 10 mice/genotype. *** P = 0.0001; Student’s t-test, two-sided. Data are mean values ± SEM. d Percentage of mice ( c ) with no lesions (None), pancreatic intraepithelial neoplasia (PanIN) only or the highest scored grade of differentiated ductal carcinoma (well, moderate, poor). e Representative photomicrographs (among 20 mice per genotype) showing H&E-stained whole pancreas cross-sections from 20wo mice (upper images). Magnified areas (squares in upper images) demonstrate normal pancreas architecture in WT and KPC:KO mice, and a carcinoma lesion in KPC mice (lower images). Scale bars = 3 mm (upper images) and 100 μm (lower images). f Representative photomicrographs of 20wo KPC and KPC:KO pancreas tissues immunostained for cleaved (p10) Caspase-1. Scale bars = 50 μm. The graph shows quantification of cleaved Caspase-1-positive epithelial staining. n = 6/group. **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. g Images of ASC (green) or cleaved (p10) Caspase-1 (red) single immunofluorescence on 20wo KPC (upper images) and KPC:KO (lower images) mouse pancreas cross-sections (representing 5 mice per genotype). Also shown (far right) are corresponding merged images (DAPI nuclear staining, blue) demonstrating cytoplasmic (yellow arrows) and nuclear (white arrows) colocalization of cleaved Caspase-1 and ASC in ductal epithelium of KPC mice. Scale bars = 10 μm. Immunoblots of ( h ) cleaved (p20) and pro (p45) Caspase-1, and ( j ) full-length (FL) and N-terminal cleaved (NT, 30kD) GSDMD, and Tubulin, on 20wo mouse pancreas tissue lysates. Each lane represents an individual mouse sample. Blots represent at least 3 independent experiments. i Representative photomicrographs of 20wo pancreas tissues immunostained for total GSDMD. Scale bars = 100 μm. The graph shows quantification of GSDMD-positive immunohistochemical epithelial staining. n = 5/group. *** P = 0.0002, **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. k ELISAs for total IL-18 and IL-1β protein levels in 20wo mouse serum. n = 5/genotype. **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. l ELISA for mature (p18) IL-18 protein levels in pancreas tissue lysates from 20wo mice. n = 5/genotype. ** P = 0.007 (WT vs KPC), 0.0067 (KPC vs KPC:KO); One-way ANOVA. Data are mean values ± SEM.

Journal: Nature Communications

Article Title: Cancer cell-intrinsic inflammasome protein ASC links innate immunity with mitochondrial metabolism in driving pancreatic cancer

doi: 10.1038/s41467-026-69398-w

Figure Lengend Snippet: a Representative 20-week-old (wo) wild-type (WT), KPC and KPC:ASC KO (KPC:KO) mouse pancreas images. Scale bars = 1 cm. b Total mass (grams; g) of pancreas in mice. n = 20 mice/group. **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. c Percentage area of whole pancreas (H&E-stained whole slide scans) containing lesions. n = 10 mice/genotype. *** P = 0.0001; Student’s t-test, two-sided. Data are mean values ± SEM. d Percentage of mice ( c ) with no lesions (None), pancreatic intraepithelial neoplasia (PanIN) only or the highest scored grade of differentiated ductal carcinoma (well, moderate, poor). e Representative photomicrographs (among 20 mice per genotype) showing H&E-stained whole pancreas cross-sections from 20wo mice (upper images). Magnified areas (squares in upper images) demonstrate normal pancreas architecture in WT and KPC:KO mice, and a carcinoma lesion in KPC mice (lower images). Scale bars = 3 mm (upper images) and 100 μm (lower images). f Representative photomicrographs of 20wo KPC and KPC:KO pancreas tissues immunostained for cleaved (p10) Caspase-1. Scale bars = 50 μm. The graph shows quantification of cleaved Caspase-1-positive epithelial staining. n = 6/group. **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. g Images of ASC (green) or cleaved (p10) Caspase-1 (red) single immunofluorescence on 20wo KPC (upper images) and KPC:KO (lower images) mouse pancreas cross-sections (representing 5 mice per genotype). Also shown (far right) are corresponding merged images (DAPI nuclear staining, blue) demonstrating cytoplasmic (yellow arrows) and nuclear (white arrows) colocalization of cleaved Caspase-1 and ASC in ductal epithelium of KPC mice. Scale bars = 10 μm. Immunoblots of ( h ) cleaved (p20) and pro (p45) Caspase-1, and ( j ) full-length (FL) and N-terminal cleaved (NT, 30kD) GSDMD, and Tubulin, on 20wo mouse pancreas tissue lysates. Each lane represents an individual mouse sample. Blots represent at least 3 independent experiments. i Representative photomicrographs of 20wo pancreas tissues immunostained for total GSDMD. Scale bars = 100 μm. The graph shows quantification of GSDMD-positive immunohistochemical epithelial staining. n = 5/group. *** P = 0.0002, **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. k ELISAs for total IL-18 and IL-1β protein levels in 20wo mouse serum. n = 5/genotype. **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM. l ELISA for mature (p18) IL-18 protein levels in pancreas tissue lysates from 20wo mice. n = 5/genotype. ** P = 0.007 (WT vs KPC), 0.0067 (KPC vs KPC:KO); One-way ANOVA. Data are mean values ± SEM.

Techniques: Staining, Immunofluorescence, Western Blot, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay

Representative photomicrographs of pancreas tissues (representing 5 mice per genotype) from a 10-week-old (wo) KPC mouse with conditionally targeted ASC alleles (KPC:fl/fl), and a parental KPC mouse, immunostained for ( a ) ASC and ( b ) cleaved (p10) Caspase-1. Scale bars = 100 μm. Magnified images depict lower staining of ASC and cleaved Caspase-1 in the ductal epithelium of KPC:fl/fl versus KPC mice. c Representative images of DAPI (nuclear; blue), ASC (green), or cleaved (p10) Caspase-1 (red) immunofluorescence on mouse pancreas cross-sections (representing 3 mice per genotype). The corresponding merged images show colocalization (white arrows) of ASC and cleaved Caspase-1 in KPC ductal lesions. Scale bars = 10 μm. d Representative pancreas images of a 10wo KPC and KPC:fl/fl mouse. Scale bars = 1 cm. e Total mass (grams; g) of pancreas in 10wo mice. n = 10 mice/group. * P = 0.016 (KPC v KPC:fl/fl), ** P = 0.0038 (WT v KPC); One-way ANOVA. Data are mean values ± SEM. f Left images depict representative low-power photomicrographs of H&E-stained whole mouse pancreas. The right images depict magnified areas (white squares in left images) of a large carcinoma lesion with poor differentiated ducts (black arrows) in KPC mice, and a small lesion with well-differentiated ducts (blue arrows) adjacent to normal acinar cells (yellow asterisk) in KPC:fl/fl mice. Scale bars = 1 mm (left images) and 100 μm (right images). g Percentage area of whole pancreas (H&E-stained whole slide scans) containing lesions. n = 10 mice/genotype. * P = 0.0207; Student’s t-test, two-sided. Data are mean values ± SEM. h Percentage of mice ( g ) with pancreatic intraepithelial neoplasia (PanIN) only or the highest grade of scored differentiated ductal carcinomas (well, moderate, poor). Representative photomicrographs of mouse pancreas cross-sections stained with antibodies against ( i ) Ki67 and ( j ) CD45. Scale bars = 100 μm. Magnified images (insets) depict reduced staining of Ki67 in the ductal epithelium of KPC:fl/fl versus KPC mice. Graphs show quantification of respective immunohistochemical staining (including age-matched WT mice) as ( i ) percentage of Ki67-positive versus total epithelial cells within ducts, and ( j ) number of total CD45-positive cells, per high power field (HPF). n = 6/group. *** P = 0.0003, **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM.

Journal: Nature Communications

Article Title: Cancer cell-intrinsic inflammasome protein ASC links innate immunity with mitochondrial metabolism in driving pancreatic cancer

doi: 10.1038/s41467-026-69398-w

Figure Lengend Snippet: Representative photomicrographs of pancreas tissues (representing 5 mice per genotype) from a 10-week-old (wo) KPC mouse with conditionally targeted ASC alleles (KPC:fl/fl), and a parental KPC mouse, immunostained for ( a ) ASC and ( b ) cleaved (p10) Caspase-1. Scale bars = 100 μm. Magnified images depict lower staining of ASC and cleaved Caspase-1 in the ductal epithelium of KPC:fl/fl versus KPC mice. c Representative images of DAPI (nuclear; blue), ASC (green), or cleaved (p10) Caspase-1 (red) immunofluorescence on mouse pancreas cross-sections (representing 3 mice per genotype). The corresponding merged images show colocalization (white arrows) of ASC and cleaved Caspase-1 in KPC ductal lesions. Scale bars = 10 μm. d Representative pancreas images of a 10wo KPC and KPC:fl/fl mouse. Scale bars = 1 cm. e Total mass (grams; g) of pancreas in 10wo mice. n = 10 mice/group. * P = 0.016 (KPC v KPC:fl/fl), ** P = 0.0038 (WT v KPC); One-way ANOVA. Data are mean values ± SEM. f Left images depict representative low-power photomicrographs of H&E-stained whole mouse pancreas. The right images depict magnified areas (white squares in left images) of a large carcinoma lesion with poor differentiated ducts (black arrows) in KPC mice, and a small lesion with well-differentiated ducts (blue arrows) adjacent to normal acinar cells (yellow asterisk) in KPC:fl/fl mice. Scale bars = 1 mm (left images) and 100 μm (right images). g Percentage area of whole pancreas (H&E-stained whole slide scans) containing lesions. n = 10 mice/genotype. * P = 0.0207; Student’s t-test, two-sided. Data are mean values ± SEM. h Percentage of mice ( g ) with pancreatic intraepithelial neoplasia (PanIN) only or the highest grade of scored differentiated ductal carcinomas (well, moderate, poor). Representative photomicrographs of mouse pancreas cross-sections stained with antibodies against ( i ) Ki67 and ( j ) CD45. Scale bars = 100 μm. Magnified images (insets) depict reduced staining of Ki67 in the ductal epithelium of KPC:fl/fl versus KPC mice. Graphs show quantification of respective immunohistochemical staining (including age-matched WT mice) as ( i ) percentage of Ki67-positive versus total epithelial cells within ducts, and ( j ) number of total CD45-positive cells, per high power field (HPF). n = 6/group. *** P = 0.0003, **** P < 0.0001; One-way ANOVA. Data are mean values ± SEM.

Techniques: Staining, Immunofluorescence, Immunohistochemical staining

a Immunoblots of pro (p45) and mature (p20) Caspase-1, and Tubulin, on pancreas tissue lysates from 10wo KPC mice treated with VHH mASC or vehicle for 24 hours. Each lane represents an individual mouse. b Mass (grams; g) of pancreas in 14wo KPC mice treated over 8 weeks with VHH mASC ( n = 7) or vehicle ( n = 5). * P = 0.0202; Student’s t-test, two-sided. Data are mean values ± SEM. c Representative photomicrographs of H&E-stained pancreas cross-sections of 14wo KPC mice treated over 8 weeks with VHH mASC or vehicle. In vehicle-treated mice poor differentiated ducts are indicated (black arrows), and in VHH mASC -treated mice moderate differentiated ducts are depicted (red arrows) along with acinar cells (yellow asterisk). Scale bars = 1 mm (whole pancreas images) and 50μm (magnified images). d Percentage of mice per treatment group ( b and c panels) with pancreatic intraepithelial neoplasia (PanIN), grades of differentiated ductal carcinomas (moderate, poor), or are lesion-free (None). e Percentage area of whole pancreas (from H&E-stained slide scans) containing lesions from 14wo VHH mASC -treated ( n = 7) versus vehicle-treated ( n = 5) KPC mice. * P = 0.040; Student’s t-test, two-sided. Data are mean values ± SEM. Representative photomicrographs of pancreas cross-sections of VHH mASC -treated versus vehicle-treated KPC mice over 8 weeks stained with antibodies against ( f ) Ki67, ( h ) cleaved (p10) Caspase-1, ( k ) CD45 and ( m ) CD3. Scale bars = 100 μm ( f ), 25μm ( h ), and 50 μm ( k , m ). j Representative confocal imaging of ASC (green) or cleaved (p10) Caspase-1 (red) immunofluorescence in pancreas from mice receiving the indicated treatment. Merged images for ASC and cleaved Caspase-1, plus DAPI nuclear staining (blue), are shown. White (nuclear) and yellow (cytosolic) arrows indicate colocalization of ASC with cleaved Caspase-1 as ASC specks. Scale bars = 10 μm. Quantification of the respective immunohistochemical staining in mouse pancreas as positive cells per high power field (HPF). n = 5/group. * P = 0.040 ( l ), ** P = 0.0033 ( g ), 0.0020 ( i ), 0.0020 ( n ); Student’s t-test, two-sided. Data are mean values ± SEM.

Journal: Nature Communications

Article Title: Cancer cell-intrinsic inflammasome protein ASC links innate immunity with mitochondrial metabolism in driving pancreatic cancer

doi: 10.1038/s41467-026-69398-w

Figure Lengend Snippet: a Immunoblots of pro (p45) and mature (p20) Caspase-1, and Tubulin, on pancreas tissue lysates from 10wo KPC mice treated with VHH mASC or vehicle for 24 hours. Each lane represents an individual mouse. b Mass (grams; g) of pancreas in 14wo KPC mice treated over 8 weeks with VHH mASC ( n = 7) or vehicle ( n = 5). * P = 0.0202; Student’s t-test, two-sided. Data are mean values ± SEM. c Representative photomicrographs of H&E-stained pancreas cross-sections of 14wo KPC mice treated over 8 weeks with VHH mASC or vehicle. In vehicle-treated mice poor differentiated ducts are indicated (black arrows), and in VHH mASC -treated mice moderate differentiated ducts are depicted (red arrows) along with acinar cells (yellow asterisk). Scale bars = 1 mm (whole pancreas images) and 50μm (magnified images). d Percentage of mice per treatment group ( b and c panels) with pancreatic intraepithelial neoplasia (PanIN), grades of differentiated ductal carcinomas (moderate, poor), or are lesion-free (None). e Percentage area of whole pancreas (from H&E-stained slide scans) containing lesions from 14wo VHH mASC -treated ( n = 7) versus vehicle-treated ( n = 5) KPC mice. * P = 0.040; Student’s t-test, two-sided. Data are mean values ± SEM. Representative photomicrographs of pancreas cross-sections of VHH mASC -treated versus vehicle-treated KPC mice over 8 weeks stained with antibodies against ( f ) Ki67, ( h ) cleaved (p10) Caspase-1, ( k ) CD45 and ( m ) CD3. Scale bars = 100 μm ( f ), 25μm ( h ), and 50 μm ( k , m ). j Representative confocal imaging of ASC (green) or cleaved (p10) Caspase-1 (red) immunofluorescence in pancreas from mice receiving the indicated treatment. Merged images for ASC and cleaved Caspase-1, plus DAPI nuclear staining (blue), are shown. White (nuclear) and yellow (cytosolic) arrows indicate colocalization of ASC with cleaved Caspase-1 as ASC specks. Scale bars = 10 μm. Quantification of the respective immunohistochemical staining in mouse pancreas as positive cells per high power field (HPF). n = 5/group. * P = 0.040 ( l ), ** P = 0.0033 ( g ), 0.0020 ( i ), 0.0020 ( n ); Student’s t-test, two-sided. Data are mean values ± SEM.

Techniques: Western Blot, Staining, Imaging, Immunofluorescence, Immunohistochemical staining

a Immunoblots for representative proteins of the five mitochondrial OXPHOS complexes (I-V) in 20-week-old (wo) wild-type (WT), KPC and KPC:ASC KO mouse pancreas tissue lysates. The samples are derived from the same experiment, but separate gel blots were processed in parallel because of different secondary conjugated antibodies used (TrueBlot DyLight 680, top blot; Alexa Fluor 680, bottom blot) to avoid interference (top blot) from Ig heavy-chain band that overlaps with ATP5A (~50kD). Arrow (bottom blot) depicts non-specific mouse Ig light-chain band. Shown on left hand-side of blots is size of molecular weight markers (kD). Each lane represents an individual mouse sample. Blots represent at least 3 independent experiments. b Quantification of mitochondrial OXPHOS complex proteins in blots ( a ) relative to corresponding total protein levels. n = 4/genotype. * P = 0.0278 (UQCRC2, KPC v KPC:KO), 0.0391 (ATP5A, KPC v KPC:KO); ** P = 0.0018 (NDUFB8, WT v KPC), 0.0088 (ATP5A, WT v KPC); *** P = 0.001 (UQCRC2, WT v KPC); One-way ANOVA. Data are mean values ± SEM. c qPCR of mtDNA copy number in pancreas tissues from 10wo and 20wo mice. n = 6/genotype. * P = 0.0424 (10wo KPC v KPC:KO); ** P = 0.0043 (10wo WT v KPC), 0.0096 (20wo WT v KPC), 0.0040 (20wo KPC v KPC:KO); One-way ANOVA. Data are mean values ± SEM. d Representative OxyBlot of protein carbonylation (oxidization) levels on pancreas protein lysates from 20wo mice. Each lane represents an individual mouse sample. e Densitometric quantification of total protein carbonylation normalized against corresponding total protein levels. n = 4/genotype. * P = 0.0175, ** P = 0.0100; One-way ANOVA. Data are mean values ± SEM. f Photomicrographs of 20wo mouse pancreas tissues (representing 5 mice per genotype) immunostained for 4-hydroxy-2-nonenal (4-HNE). Scale bars = 100 μm. g Lactate levels in 10wo and 20wo mouse pancreas tissue lysates. n = 5/genotype. * P = 0.0350 (10wo KPC v KPC:KO), 0.0231 (20wo WT v KPC); *** P = 0.006 (10wo KPC v KPC:KO); One-way ANOVA. Data are mean values ± SEM. h Representative Opal multiplex immunofluorescence confocal images of single staining for DAPI (nuclear; blue), ASC (violet), cleaved (p10) Caspase-1 (blue-green), UQCRC2 (orange), NDUFB8 (red) and either CK19 or CD45 (green), along with merged images, in serial pancreas cross-sections from 20wo KPC mice (representative of 3 mice). Dotted white squares in merged images depict magnified areas in Supplementary Fig. . Scale bars = 100 μm. i Computationally reconstructed cellular spatial maps of representative merged immunofluorescence images (panel h ) showing colocalization of inflammasome (ASC, cleaved Caspase-1) and/or mitochondrial (NDUFB8, UQCRC2) markers with CK19-positive ductal epithelial cells or CD45-positive immune cells. Also shown is quantification of the composition (percentage) of inflammasome and/or mitochondrial markers colocalized within the ductal epithelial or immune cell populations.

Journal: Nature Communications

Article Title: Cancer cell-intrinsic inflammasome protein ASC links innate immunity with mitochondrial metabolism in driving pancreatic cancer

doi: 10.1038/s41467-026-69398-w

Figure Lengend Snippet: a Immunoblots for representative proteins of the five mitochondrial OXPHOS complexes (I-V) in 20-week-old (wo) wild-type (WT), KPC and KPC:ASC KO mouse pancreas tissue lysates. The samples are derived from the same experiment, but separate gel blots were processed in parallel because of different secondary conjugated antibodies used (TrueBlot DyLight 680, top blot; Alexa Fluor 680, bottom blot) to avoid interference (top blot) from Ig heavy-chain band that overlaps with ATP5A (~50kD). Arrow (bottom blot) depicts non-specific mouse Ig light-chain band. Shown on left hand-side of blots is size of molecular weight markers (kD). Each lane represents an individual mouse sample. Blots represent at least 3 independent experiments. b Quantification of mitochondrial OXPHOS complex proteins in blots ( a ) relative to corresponding total protein levels. n = 4/genotype. * P = 0.0278 (UQCRC2, KPC v KPC:KO), 0.0391 (ATP5A, KPC v KPC:KO); ** P = 0.0018 (NDUFB8, WT v KPC), 0.0088 (ATP5A, WT v KPC); *** P = 0.001 (UQCRC2, WT v KPC); One-way ANOVA. Data are mean values ± SEM. c qPCR of mtDNA copy number in pancreas tissues from 10wo and 20wo mice. n = 6/genotype. * P = 0.0424 (10wo KPC v KPC:KO); ** P = 0.0043 (10wo WT v KPC), 0.0096 (20wo WT v KPC), 0.0040 (20wo KPC v KPC:KO); One-way ANOVA. Data are mean values ± SEM. d Representative OxyBlot of protein carbonylation (oxidization) levels on pancreas protein lysates from 20wo mice. Each lane represents an individual mouse sample. e Densitometric quantification of total protein carbonylation normalized against corresponding total protein levels. n = 4/genotype. * P = 0.0175, ** P = 0.0100; One-way ANOVA. Data are mean values ± SEM. f Photomicrographs of 20wo mouse pancreas tissues (representing 5 mice per genotype) immunostained for 4-hydroxy-2-nonenal (4-HNE). Scale bars = 100 μm. g Lactate levels in 10wo and 20wo mouse pancreas tissue lysates. n = 5/genotype. * P = 0.0350 (10wo KPC v KPC:KO), 0.0231 (20wo WT v KPC); *** P = 0.006 (10wo KPC v KPC:KO); One-way ANOVA. Data are mean values ± SEM. h Representative Opal multiplex immunofluorescence confocal images of single staining for DAPI (nuclear; blue), ASC (violet), cleaved (p10) Caspase-1 (blue-green), UQCRC2 (orange), NDUFB8 (red) and either CK19 or CD45 (green), along with merged images, in serial pancreas cross-sections from 20wo KPC mice (representative of 3 mice). Dotted white squares in merged images depict magnified areas in Supplementary Fig. . Scale bars = 100 μm. i Computationally reconstructed cellular spatial maps of representative merged immunofluorescence images (panel h ) showing colocalization of inflammasome (ASC, cleaved Caspase-1) and/or mitochondrial (NDUFB8, UQCRC2) markers with CK19-positive ductal epithelial cells or CD45-positive immune cells. Also shown is quantification of the composition (percentage) of inflammasome and/or mitochondrial markers colocalized within the ductal epithelial or immune cell populations.

Techniques: Western Blot, Derivative Assay, Molecular Weight, Multiplex Assay, Immunofluorescence, Staining

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