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    Cell Signaling Technology Inc rnase a
    Rnase A, 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 rnase a
    Rnase A, 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 rnase a
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    Cell Signaling Technology Inc mouse il 1β
    WT161 decreased the levels of the pro-inflammatory factors in the colonic tissue of DSS-induced colitis mice and activated peritoneal macrophages induced by LPS + DSS. (A,B) The colonic explants isolated from mice in the animal experiments were cultured for 24 h, and the <t>interleukin</t> <t>(IL)-1β</t> (A) and IL-6 (B) levels in the supernatant was detected by ELISA. For comparison, we extracted the same weight of colon explants (DMSO + H 2 O group, n = 3; other groups, n = 7). (C,D) The IL-1β (C) and IL-6 (D) levels in the supernatant culture of LPS + DSS-activated peritoneal macrophages. For (A–D) , the p -values were calculated using one-way ANOVA with the Bonferroni’s test. ** p < 0.01; **** p < 0.0001. DSS, dextran sulfate sodium; DMSO, dimethyl sulfoxide.
    Mouse Il 1β, 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 "Identification of WT161 as a Potent Agent for the Treatment of Colitis by Targeting the Nucleotide-Binding Domain-Like Receptor Family Pyrin Domain Containing 3 Inflammasome"

    Article Title: Identification of WT161 as a Potent Agent for the Treatment of Colitis by Targeting the Nucleotide-Binding Domain-Like Receptor Family Pyrin Domain Containing 3 Inflammasome

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2022.780179

    WT161 decreased the levels of the pro-inflammatory factors in the colonic tissue of DSS-induced colitis mice and activated peritoneal macrophages induced by LPS + DSS. (A,B) The colonic explants isolated from mice in the animal experiments were cultured for 24 h, and the interleukin (IL)-1β (A) and IL-6 (B) levels in the supernatant was detected by ELISA. For comparison, we extracted the same weight of colon explants (DMSO + H 2 O group, n = 3; other groups, n = 7). (C,D) The IL-1β (C) and IL-6 (D) levels in the supernatant culture of LPS + DSS-activated peritoneal macrophages. For (A–D) , the p -values were calculated using one-way ANOVA with the Bonferroni’s test. ** p < 0.01; **** p < 0.0001. DSS, dextran sulfate sodium; DMSO, dimethyl sulfoxide.
    Figure Legend Snippet: WT161 decreased the levels of the pro-inflammatory factors in the colonic tissue of DSS-induced colitis mice and activated peritoneal macrophages induced by LPS + DSS. (A,B) The colonic explants isolated from mice in the animal experiments were cultured for 24 h, and the interleukin (IL)-1β (A) and IL-6 (B) levels in the supernatant was detected by ELISA. For comparison, we extracted the same weight of colon explants (DMSO + H 2 O group, n = 3; other groups, n = 7). (C,D) The IL-1β (C) and IL-6 (D) levels in the supernatant culture of LPS + DSS-activated peritoneal macrophages. For (A–D) , the p -values were calculated using one-way ANOVA with the Bonferroni’s test. ** p < 0.01; **** p < 0.0001. DSS, dextran sulfate sodium; DMSO, dimethyl sulfoxide.

    Techniques Used: Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

    WT161 decreased NLRP3 inflammasome activation in the peritoneal macrophages. (A) Mouse peritoneal macrophages were pre-treated with WT161, and then primed with lipopolysaccharide (LPS), followed by stimulation with nigericin. The interleukin (IL)-1β in the supernatant was detected by ELISA. (B,C) The level of cleaved caspase-1 (p10) and mature IL-1β (p17) upon nigericin stimulation in the supernatant (SN) by WT161 treatment was analyzed by western blot (B) and quantitative analysis (C) . (D) Immunoblot analysis of ASC oligomerization in lysates of LPS-primed peritoneal macrophages treated with WT161 and then stimulated with nigericin. (E) Quantitative analysis of ASC oligomerization. (F) Immunofluorescence microscopy analysis of ASC Speck (arrow) in LPS-primed peritoneal macrophages treated with or without WT161, and then stimulated with ATP or nigericin. Scale bars: 10 μm. (G) Percentage of cells with ASC specks. The image and data shown in (A–F) are representative of three independent experiments. For (A,C,E,G) , the p -values were calculated using two-way ANOVA with the Bonferroni’s multiple comparisons test. * p < 0.05; **** p < 0.0001; while p > 0.05 displayed as ns. DMSO, dimethyl sulfoxide; LN, LPS + nigericin; SN, supernatant; MOCK, blank control.
    Figure Legend Snippet: WT161 decreased NLRP3 inflammasome activation in the peritoneal macrophages. (A) Mouse peritoneal macrophages were pre-treated with WT161, and then primed with lipopolysaccharide (LPS), followed by stimulation with nigericin. The interleukin (IL)-1β in the supernatant was detected by ELISA. (B,C) The level of cleaved caspase-1 (p10) and mature IL-1β (p17) upon nigericin stimulation in the supernatant (SN) by WT161 treatment was analyzed by western blot (B) and quantitative analysis (C) . (D) Immunoblot analysis of ASC oligomerization in lysates of LPS-primed peritoneal macrophages treated with WT161 and then stimulated with nigericin. (E) Quantitative analysis of ASC oligomerization. (F) Immunofluorescence microscopy analysis of ASC Speck (arrow) in LPS-primed peritoneal macrophages treated with or without WT161, and then stimulated with ATP or nigericin. Scale bars: 10 μm. (G) Percentage of cells with ASC specks. The image and data shown in (A–F) are representative of three independent experiments. For (A,C,E,G) , the p -values were calculated using two-way ANOVA with the Bonferroni’s multiple comparisons test. * p < 0.05; **** p < 0.0001; while p > 0.05 displayed as ns. DMSO, dimethyl sulfoxide; LN, LPS + nigericin; SN, supernatant; MOCK, blank control.

    Techniques Used: Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Immunofluorescence, Microscopy

    rnase a  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc mouse il 1β
    C646 suppresses NLRP3 inflammasome activity in the DSS-induced colitis model. (A) Immunoblot analysis of cleaved caspase 1 and NLRP3. Data were normalized to the expression of β-actin as reference. (B) and (C) Protein level of the <t>cleaved</t> <t>IL-1β</t> and IL-6 in colon homogenates was determined by ELISA. (D) Immunoblot analysis of ASC oligomerization in cross-linked cytosolic pellets of DSS-induced colitis treated with C646 or DMSO. The results of (B) and (C) were expressed as mean ± SD. Statistics were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.
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    1) Product Images from "C646 Protects Against DSS-Induced Colitis Model by Targeting NLRP3 Inflammasome"

    Article Title: C646 Protects Against DSS-Induced Colitis Model by Targeting NLRP3 Inflammasome

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2021.707610

    C646 suppresses NLRP3 inflammasome activity in the DSS-induced colitis model. (A) Immunoblot analysis of cleaved caspase 1 and NLRP3. Data were normalized to the expression of β-actin as reference. (B) and (C) Protein level of the cleaved IL-1β and IL-6 in colon homogenates was determined by ELISA. (D) Immunoblot analysis of ASC oligomerization in cross-linked cytosolic pellets of DSS-induced colitis treated with C646 or DMSO. The results of (B) and (C) were expressed as mean ± SD. Statistics were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.
    Figure Legend Snippet: C646 suppresses NLRP3 inflammasome activity in the DSS-induced colitis model. (A) Immunoblot analysis of cleaved caspase 1 and NLRP3. Data were normalized to the expression of β-actin as reference. (B) and (C) Protein level of the cleaved IL-1β and IL-6 in colon homogenates was determined by ELISA. (D) Immunoblot analysis of ASC oligomerization in cross-linked cytosolic pellets of DSS-induced colitis treated with C646 or DMSO. The results of (B) and (C) were expressed as mean ± SD. Statistics were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.

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

    C646 suppresses the NLRP3 inflammasome at the priming and assembly steps. (A–D) Mouse peritoneal macrophages were treated with C646 and then primed with LPS, followed by stimulation with nigericin. The supernatant (SN) was subjected to ELISA assay for IL-1β, IL-6, TNF-α, and LDH. (E–H) Mouse peritoneal macrophages were primed with LPS and then treated with C646, followed by stimulation with nigericin. The supernatant (SN) was subjected to ELISA assay for IL-1β, IL-6, TNF-α, and LDH. For (A) to (H), values are mean ± SD. Statistics were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.
    Figure Legend Snippet: C646 suppresses the NLRP3 inflammasome at the priming and assembly steps. (A–D) Mouse peritoneal macrophages were treated with C646 and then primed with LPS, followed by stimulation with nigericin. The supernatant (SN) was subjected to ELISA assay for IL-1β, IL-6, TNF-α, and LDH. (E–H) Mouse peritoneal macrophages were primed with LPS and then treated with C646, followed by stimulation with nigericin. The supernatant (SN) was subjected to ELISA assay for IL-1β, IL-6, TNF-α, and LDH. For (A) to (H), values are mean ± SD. Statistics were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    C646 suppresses the NLRP3 inflammasome activated by different agonists. (A–H) ELISA assay for IL-1β, IL-6, TNF-α, and LDH in the supernatant of mouse peritoneal macrophages that were treated with C646 or DMSO upon stimulation of LPS + ATP or LPS + MSU. (I) and (J) Mouse peritoneal macrophages were primed with LPS and then treated with C646, followed by stimulation with ATP or MSU. The supernatant (SN) was subjected to Western blot analysis for the indicated protein levels. Data are representative of three independent experiments. For (A) to (H), values are mean ± SD. Statistics were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.
    Figure Legend Snippet: C646 suppresses the NLRP3 inflammasome activated by different agonists. (A–H) ELISA assay for IL-1β, IL-6, TNF-α, and LDH in the supernatant of mouse peritoneal macrophages that were treated with C646 or DMSO upon stimulation of LPS + ATP or LPS + MSU. (I) and (J) Mouse peritoneal macrophages were primed with LPS and then treated with C646, followed by stimulation with ATP or MSU. The supernatant (SN) was subjected to Western blot analysis for the indicated protein levels. Data are representative of three independent experiments. For (A) to (H), values are mean ± SD. Statistics were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Western Blot

    C646 inhibits the activation of the NF-κB signaling. (A–D) Quantitative PCR analysis of mRNA expression of TNF-α, IL-6, IL-1β, and NLRP3 in primary peritoneal macrophages treated with C646 and primed with LPS for the indicated hours. (E) Immunoblot analysis of p-p65, p65, NLRP3, and IL-1β in mouse peritoneal macrophages treated with C646 and then primed with LPS for the indicated hours. Values are mean ± SD. For (A) to (D), values were analyzed using two-way ANOVA and the Bonferroni test. * p < 0.05, ** p < 0.01 and *** p < 0.001.
    Figure Legend Snippet: C646 inhibits the activation of the NF-κB signaling. (A–D) Quantitative PCR analysis of mRNA expression of TNF-α, IL-6, IL-1β, and NLRP3 in primary peritoneal macrophages treated with C646 and primed with LPS for the indicated hours. (E) Immunoblot analysis of p-p65, p65, NLRP3, and IL-1β in mouse peritoneal macrophages treated with C646 and then primed with LPS for the indicated hours. Values are mean ± SD. For (A) to (D), values were analyzed using two-way ANOVA and the Bonferroni test. * p < 0.05, ** p < 0.01 and *** p < 0.001.

    Techniques Used: Activation Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot

    C646 ameliorates LPS-induced acute systemic inflammation in vivo. C57BL/6 mice were given intraperitoneal (i.p.) injection of N.S or C646 (10 or 20 mg/kg) 30 min before i. p. injection of LPS (20 mg/kg) for 8 h (n = 3–5/group). ELISA of serum IL-1β (A) , IL-6 (B) , and TNF-α (C) . Representative H&E images of lung sections (D) were collected. Scale bar, 100 μm. For (A) to (C), values were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.
    Figure Legend Snippet: C646 ameliorates LPS-induced acute systemic inflammation in vivo. C57BL/6 mice were given intraperitoneal (i.p.) injection of N.S or C646 (10 or 20 mg/kg) 30 min before i. p. injection of LPS (20 mg/kg) for 8 h (n = 3–5/group). ELISA of serum IL-1β (A) , IL-6 (B) , and TNF-α (C) . Representative H&E images of lung sections (D) were collected. Scale bar, 100 μm. For (A) to (C), values were analyzed using one-way ANOVA and the Bonferroni test. ** p < 0.05; *** p < 0.01; **** p < 0.0001.

    Techniques Used: In Vivo, Injection, Enzyme-linked Immunosorbent Assay

    rnase a  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc mouse il 1β
    WT161 decreased the levels of the pro-inflammatory factors in the colonic tissue of DSS-induced colitis mice and activated peritoneal macrophages induced by LPS + DSS. (A,B) The colonic explants isolated from mice in the animal experiments were cultured for 24 h, and the <t>interleukin</t> <t>(IL)-1β</t> (A) and IL-6 (B) levels in the supernatant was detected by ELISA. For comparison, we extracted the same weight of colon explants (DMSO + H 2 O group, n = 3; other groups, n = 7). (C,D) The IL-1β (C) and IL-6 (D) levels in the supernatant culture of LPS + DSS-activated peritoneal macrophages. For (A–D) , the p -values were calculated using one-way ANOVA with the Bonferroni’s test. ** p < 0.01; **** p < 0.0001. DSS, dextran sulfate sodium; DMSO, dimethyl sulfoxide.
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    WT161 decreased the levels of the pro-inflammatory factors in the colonic tissue of DSS-induced colitis mice and activated peritoneal macrophages induced by LPS + DSS. (A,B) The colonic explants isolated from mice in the animal experiments were cultured for 24 h, and the interleukin (IL)-1β (A) and IL-6 (B) levels in the supernatant was detected by ELISA. For comparison, we extracted the same weight of colon explants (DMSO + H 2 O group, n = 3; other groups, n = 7). (C,D) The IL-1β (C) and IL-6 (D) levels in the supernatant culture of LPS + DSS-activated peritoneal macrophages. For (A–D) , the p -values were calculated using one-way ANOVA with the Bonferroni’s test. ** p < 0.01; **** p < 0.0001. DSS, dextran sulfate sodium; DMSO, dimethyl sulfoxide.

    Journal: Frontiers in Pharmacology

    Article Title: Identification of WT161 as a Potent Agent for the Treatment of Colitis by Targeting the Nucleotide-Binding Domain-Like Receptor Family Pyrin Domain Containing 3 Inflammasome

    doi: 10.3389/fphar.2022.780179

    Figure Lengend Snippet: WT161 decreased the levels of the pro-inflammatory factors in the colonic tissue of DSS-induced colitis mice and activated peritoneal macrophages induced by LPS + DSS. (A,B) The colonic explants isolated from mice in the animal experiments were cultured for 24 h, and the interleukin (IL)-1β (A) and IL-6 (B) levels in the supernatant was detected by ELISA. For comparison, we extracted the same weight of colon explants (DMSO + H 2 O group, n = 3; other groups, n = 7). (C,D) The IL-1β (C) and IL-6 (D) levels in the supernatant culture of LPS + DSS-activated peritoneal macrophages. For (A–D) , the p -values were calculated using one-way ANOVA with the Bonferroni’s test. ** p < 0.01; **** p < 0.0001. DSS, dextran sulfate sodium; DMSO, dimethyl sulfoxide.

    Article Snippet: WT161 (cat. no. 1206731-57-8) from Selleck Chemicals; DSS (cat. no. 0216011080, 36,000–50,000 molecular weight) from MP Biomedicals; ultrapure lipopolysaccharide (LPS) ( E. coli 0111: B4, cat. no. tlrl-3pelps), ATP (cat. no. tlrl-atpl), and nigericin (cat. no. tlrl-nig), monosodium urate (MSU) (cat. no. tlrl-msu) from InvivoGen; Protein A/G PLUS-Agarose (cat. no. sc-2003) from Santa Cruz; mouse immunoglobin IgG protein (cat. no. ab198772) from Abcam, and cell lysis buffer (CLB) (cat. no. 9803) from Cell Signaling Technology; mouse IL-1β (cat. no. 88–7013), IL-6 (cat. no. 88-7064-88) ELISA kits (Thermo Fisher).

    Techniques: Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

    WT161 decreased NLRP3 inflammasome activation in the peritoneal macrophages. (A) Mouse peritoneal macrophages were pre-treated with WT161, and then primed with lipopolysaccharide (LPS), followed by stimulation with nigericin. The interleukin (IL)-1β in the supernatant was detected by ELISA. (B,C) The level of cleaved caspase-1 (p10) and mature IL-1β (p17) upon nigericin stimulation in the supernatant (SN) by WT161 treatment was analyzed by western blot (B) and quantitative analysis (C) . (D) Immunoblot analysis of ASC oligomerization in lysates of LPS-primed peritoneal macrophages treated with WT161 and then stimulated with nigericin. (E) Quantitative analysis of ASC oligomerization. (F) Immunofluorescence microscopy analysis of ASC Speck (arrow) in LPS-primed peritoneal macrophages treated with or without WT161, and then stimulated with ATP or nigericin. Scale bars: 10 μm. (G) Percentage of cells with ASC specks. The image and data shown in (A–F) are representative of three independent experiments. For (A,C,E,G) , the p -values were calculated using two-way ANOVA with the Bonferroni’s multiple comparisons test. * p < 0.05; **** p < 0.0001; while p > 0.05 displayed as ns. DMSO, dimethyl sulfoxide; LN, LPS + nigericin; SN, supernatant; MOCK, blank control.

    Journal: Frontiers in Pharmacology

    Article Title: Identification of WT161 as a Potent Agent for the Treatment of Colitis by Targeting the Nucleotide-Binding Domain-Like Receptor Family Pyrin Domain Containing 3 Inflammasome

    doi: 10.3389/fphar.2022.780179

    Figure Lengend Snippet: WT161 decreased NLRP3 inflammasome activation in the peritoneal macrophages. (A) Mouse peritoneal macrophages were pre-treated with WT161, and then primed with lipopolysaccharide (LPS), followed by stimulation with nigericin. The interleukin (IL)-1β in the supernatant was detected by ELISA. (B,C) The level of cleaved caspase-1 (p10) and mature IL-1β (p17) upon nigericin stimulation in the supernatant (SN) by WT161 treatment was analyzed by western blot (B) and quantitative analysis (C) . (D) Immunoblot analysis of ASC oligomerization in lysates of LPS-primed peritoneal macrophages treated with WT161 and then stimulated with nigericin. (E) Quantitative analysis of ASC oligomerization. (F) Immunofluorescence microscopy analysis of ASC Speck (arrow) in LPS-primed peritoneal macrophages treated with or without WT161, and then stimulated with ATP or nigericin. Scale bars: 10 μm. (G) Percentage of cells with ASC specks. The image and data shown in (A–F) are representative of three independent experiments. For (A,C,E,G) , the p -values were calculated using two-way ANOVA with the Bonferroni’s multiple comparisons test. * p < 0.05; **** p < 0.0001; while p > 0.05 displayed as ns. DMSO, dimethyl sulfoxide; LN, LPS + nigericin; SN, supernatant; MOCK, blank control.

    Article Snippet: WT161 (cat. no. 1206731-57-8) from Selleck Chemicals; DSS (cat. no. 0216011080, 36,000–50,000 molecular weight) from MP Biomedicals; ultrapure lipopolysaccharide (LPS) ( E. coli 0111: B4, cat. no. tlrl-3pelps), ATP (cat. no. tlrl-atpl), and nigericin (cat. no. tlrl-nig), monosodium urate (MSU) (cat. no. tlrl-msu) from InvivoGen; Protein A/G PLUS-Agarose (cat. no. sc-2003) from Santa Cruz; mouse immunoglobin IgG protein (cat. no. ab198772) from Abcam, and cell lysis buffer (CLB) (cat. no. 9803) from Cell Signaling Technology; mouse IL-1β (cat. no. 88–7013), IL-6 (cat. no. 88-7064-88) ELISA kits (Thermo Fisher).

    Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Immunofluorescence, Microscopy