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il 1α  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology il 1α
    Il 1α, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/il+1%CE%B1/IL-1%CE%B1/10__3390_slash_ijms27073232-271-8-21
    Average 93 stars, based on 104 article reviews
    il 1α - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Polymer:

    Article Title: Comparison of Tissue Factors in the Ontogenetic Aspects of Human Cholesteatoma
    Article Snippet: .. The HiDef DetectionTM HRP Polymer System (954D-30, Cell Marque, Rocklin, CA, USA) was used to ready the slides for IHC staining, and to identify the following markers in the tissue samples: HβD-2 (sc-20798, rabbit, working dilution 1:200, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); HβD-4 (sc-59496, mouse, working dilution 1:50, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); IL-1α (sc-9983, mouse, working dilution 1:50, Santa Cruz Biotechnology, Inc., Paso Robles, CA, USA); IL-10 (ab34843, rabbit, working dilution 1:400, Abcam, Cambridge, UK); Ki-67 (1325506A, rabbit, 1:100, Cell Marque, Rocklin, CA, USA); NF-κβ (sc-109; rabbit, 1:200 dilution, Santa Cruz Biotechnology, Inc., Paso Robles, CA, USA); vascular endothelial growth factor (VEGF) (orb191500, rabbit, 1:100, Biorbyt Ltd., Cambridge, UK); SHH (LS-C49806, rabbit, 1:100, LifeSpan BioSciences, Inc., Seattle, WA, USA); MMP2 (sc-53630, mouse, 1:100, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); MMP-9 (sc-10737, rabbit, 1:100, Santa Cruz Biotechnology, Inc., Santa Cruz, Dallas, TX, USA); TIMP2 (sc-21735, mouse, 1:100, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); and TIMP-4 (sc-30076, rabbit, 1:100, Santa Cruz Biotechnology, Inc., Santa Cruz, Dallas, TX, USA). .. Furthermore, tissue samples were rinsed in wash buffer (TRIS; T0083, Diapath S.p.A., Martinengo, Italy) two times for 5 min, then placed in a microwave oven for 20 min in boiling EDTA buffer (T0103, Diapath S.p.A., Martinengo, Italy) and cooled to 65 °C for approximately 20 min. Then, the material was placed in TRIS wash buffer and blocked with 3% peroxidase block (925B-02, Cell Marque, Rocklin, CA, USA) for 10 min. All of the antibodies used in this study were diluted with antibody diluent (938B-05, Cell Marque, Rocklin, CA, USA).

    Immunohistochemistry:

    Article Title: Comparison of Tissue Factors in the Ontogenetic Aspects of Human Cholesteatoma
    Article Snippet: .. The HiDef DetectionTM HRP Polymer System (954D-30, Cell Marque, Rocklin, CA, USA) was used to ready the slides for IHC staining, and to identify the following markers in the tissue samples: HβD-2 (sc-20798, rabbit, working dilution 1:200, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); HβD-4 (sc-59496, mouse, working dilution 1:50, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); IL-1α (sc-9983, mouse, working dilution 1:50, Santa Cruz Biotechnology, Inc., Paso Robles, CA, USA); IL-10 (ab34843, rabbit, working dilution 1:400, Abcam, Cambridge, UK); Ki-67 (1325506A, rabbit, 1:100, Cell Marque, Rocklin, CA, USA); NF-κβ (sc-109; rabbit, 1:200 dilution, Santa Cruz Biotechnology, Inc., Paso Robles, CA, USA); vascular endothelial growth factor (VEGF) (orb191500, rabbit, 1:100, Biorbyt Ltd., Cambridge, UK); SHH (LS-C49806, rabbit, 1:100, LifeSpan BioSciences, Inc., Seattle, WA, USA); MMP2 (sc-53630, mouse, 1:100, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); MMP-9 (sc-10737, rabbit, 1:100, Santa Cruz Biotechnology, Inc., Santa Cruz, Dallas, TX, USA); TIMP2 (sc-21735, mouse, 1:100, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); and TIMP-4 (sc-30076, rabbit, 1:100, Santa Cruz Biotechnology, Inc., Santa Cruz, Dallas, TX, USA). .. Furthermore, tissue samples were rinsed in wash buffer (TRIS; T0083, Diapath S.p.A., Martinengo, Italy) two times for 5 min, then placed in a microwave oven for 20 min in boiling EDTA buffer (T0103, Diapath S.p.A., Martinengo, Italy) and cooled to 65 °C for approximately 20 min. Then, the material was placed in TRIS wash buffer and blocked with 3% peroxidase block (925B-02, Cell Marque, Rocklin, CA, USA) for 10 min. All of the antibodies used in this study were diluted with antibody diluent (938B-05, Cell Marque, Rocklin, CA, USA).

    Staining:

    Article Title: Comparison of Tissue Factors in the Ontogenetic Aspects of Human Cholesteatoma
    Article Snippet: .. The HiDef DetectionTM HRP Polymer System (954D-30, Cell Marque, Rocklin, CA, USA) was used to ready the slides for IHC staining, and to identify the following markers in the tissue samples: HβD-2 (sc-20798, rabbit, working dilution 1:200, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); HβD-4 (sc-59496, mouse, working dilution 1:50, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); IL-1α (sc-9983, mouse, working dilution 1:50, Santa Cruz Biotechnology, Inc., Paso Robles, CA, USA); IL-10 (ab34843, rabbit, working dilution 1:400, Abcam, Cambridge, UK); Ki-67 (1325506A, rabbit, 1:100, Cell Marque, Rocklin, CA, USA); NF-κβ (sc-109; rabbit, 1:200 dilution, Santa Cruz Biotechnology, Inc., Paso Robles, CA, USA); vascular endothelial growth factor (VEGF) (orb191500, rabbit, 1:100, Biorbyt Ltd., Cambridge, UK); SHH (LS-C49806, rabbit, 1:100, LifeSpan BioSciences, Inc., Seattle, WA, USA); MMP2 (sc-53630, mouse, 1:100, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); MMP-9 (sc-10737, rabbit, 1:100, Santa Cruz Biotechnology, Inc., Santa Cruz, Dallas, TX, USA); TIMP2 (sc-21735, mouse, 1:100, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); and TIMP-4 (sc-30076, rabbit, 1:100, Santa Cruz Biotechnology, Inc., Santa Cruz, Dallas, TX, USA). .. Furthermore, tissue samples were rinsed in wash buffer (TRIS; T0083, Diapath S.p.A., Martinengo, Italy) two times for 5 min, then placed in a microwave oven for 20 min in boiling EDTA buffer (T0103, Diapath S.p.A., Martinengo, Italy) and cooled to 65 °C for approximately 20 min. Then, the material was placed in TRIS wash buffer and blocked with 3% peroxidase block (925B-02, Cell Marque, Rocklin, CA, USA) for 10 min. All of the antibodies used in this study were diluted with antibody diluent (938B-05, Cell Marque, Rocklin, CA, USA).

    Article Title: Ganglioside GA2-mediated caspase-11 activation drives macrophage pyroptosis aggravating intimal hyperplasia after arterial injury
    Article Snippet: .. Primary Antibody were used for immunofluorescent staining as follows: caspase-11 (1:100, MAB86481, R&D Systems); BID (1:50, sc-373939, Santa Cruz); F4/80 (1:100, 27044-1-AP, Proteintech); F4/80 (1:100, ab16911, Abcam); IL-1α (1:50, sc-9983, Santa Cruz); N-GSDME (1:100, 38821S, CST); Ganglioside GA2 (1:100, ab23942, Abcam), CD68 (1:100, ab955, Abcam). .. Secondary goat anti-mouse (FITC) antibody (1:100, SA00003-1, Proteintech), goat anti-rat (AF647) antibody (1:200, ab150159, Abcam), goat anti-rabbit (Cy3) antibody (1:200, bs-0295G-Cy3, BIOSS), goat anti-rat (Cy3) antibody (1:200, bs-0293G-Cy3, BIOSS), goat anti-rabbit (FITC) antibody (1:200, bs-0296G-FITC, BIOSS) and goat anti-mouse (AF647) antibody (bs-0296G-AF647, BIOSS) were used.

    Article Title: Ganglioside GA2-mediated caspase-11 activation drives macrophage pyroptosis aggravating intimal hyperplasia after arterial injury.
    Article Snippet: Intimal hyperplasia (IH) remains a significant clinical problem, causing vascular intervention failure.. This study aimed to elucidate whether gangliosides GA2 accumulated in atherosclerotic mouse aortae and plasma promote the development of IH.. We identified that GA2 was remarkably accumulated in both artery and plasma of atherosclerotic patients and mice.

    Lysis:

    Article Title: MK2/p38/p53 negatively regulate basal IL-1β and non-canonical NF-κB signaling
    Article Snippet: .. The next day, the cells were treated with IL-1α, Nutlin-3 (sc- 45061, Santa Cruz Biotechnology (SCBT)), zDEVDfmk (HY-12466, Hycultec), zVADfmk (4026865, Bachem), Pifithrin μ (HY-10940, Hycultec), or Pifithrin α (506132, Sigma-Aldrich) as indicated, and then lysed using kinase lysis buffer supplemented with protease inhibitors (B14001, Bimake) and phosphatase inhibitors (B15001, Bimake). ..

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages
    Article Snippet: .. The next day, the cells were treated with IL-1α, LPS, B022, BIRB 796 (Axon Medchem BV, Groningen, The Netherlands), Nutlin3 (sc-45061, SCBT, Dallas, TX, USA), zDEVDfmk (HY-12466, Hycultec GmbH, Beutelsberg, Germany), zVADfmk (4026865, Bachem Holding AG, Bubendorf BL, Switzerland), Pifithrin μ (HY-10940, Hycultec GmbH, Beutelsberg, Germany), or Pifithrin α (506132, Sigma-Aldrich, Merck, St. Louis, MO, USA) as indicated, and then lysed using kinase lysis buffer supplemented with protease inhibitors (B14001, Bimake, Houston, TX, USA) and phosphatase inhibitors (B15001, Bimake, Houston, TX, USA). ..

    Immunohistochemical staining:

    Article Title: Supplementary Information for Cryptotanshinone-loaded cerasomes formulation: In vitro drug release, in vivo pharmacokinetics and in vivo efficacy for topical therapy of acne
    Article Snippet: .. Materials Immunohistochemical reagents: IL-1α antibody, AR antibody detection kit (Rabbit anti rat, Abcam, Cambridge, UK) and DAB Kit (Boster Biological Engineering Co., Ltd, Wuhan, China),Western blot Kit: IL-1α, AR antibody (Rabbit anti rat, Millipore company, American), PVDF membrane (Millipore, MA, USA), Tris-base (Sigma), skimmed milk powder, HRP-labeled Secondary antibody (Boster Biological Engineering Co., Ltd, Wuhan, China), β-actin (Santa Cruz). ..

    Membrane:

    Article Title: Supplementary Information for Cryptotanshinone-loaded cerasomes formulation: In vitro drug release, in vivo pharmacokinetics and in vivo efficacy for topical therapy of acne
    Article Snippet: .. Materials Immunohistochemical reagents: IL-1α antibody, AR antibody detection kit (Rabbit anti rat, Abcam, Cambridge, UK) and DAB Kit (Boster Biological Engineering Co., Ltd, Wuhan, China),Western blot Kit: IL-1α, AR antibody (Rabbit anti rat, Millipore company, American), PVDF membrane (Millipore, MA, USA), Tris-base (Sigma), skimmed milk powder, HRP-labeled Secondary antibody (Boster Biological Engineering Co., Ltd, Wuhan, China), β-actin (Santa Cruz). ..

    SDS Page:

    Article Title: SARS-CoV-2 and its ORF3a, E and M viroporins activate inflammasome in human macrophages and induce of IL-1 α in pulmonary epithelial and endothelial cells
    Article Snippet: .. 10 μg of proteins were separated in 12.5% SDS – PAGE gels and blotted onto nitrocellulose or PVDF membranes by semi-wet transfer for 10 min at 2.5 A and 25 V. Membranes were blocked in TBS-T with 5% non – fat milk for 1 h in RT following incubation overnight at 4 °C with primary (1:1000) antibodies: Flag (Sigma, Cat. F1804), IL-1β (Cell Signaling Technology, MA, USA, Cat. 2022 and Affinity Biosciences, Ohio, USA, Cat. AF4012), IL-1α (Santa Cruz, CA, USA, Cat. sc-9983), NLRP3 (Adipogen, Switzerland, Cat. AG-20B-0014-C100), GSDMD (Cell Signaling Technology, Cat. 96458), β-actin (Novus, CO, USA, Cat.NB600), GAPDH (Millipore, MA, USA, Cat. MAB374; 1:5000). .. Appropriate mouse or rabbit HRP-conjugated secondary antibody (Vector Laboratories Inc., CA, USA, Cat. PI-2000, PI-1000; 1:10000) was used to detect primary antibodies.

    Article Title: SARS-CoV-2 and its ORF3a, E and M viroporins activate inflammasome in human macrophages and induce of IL-1α in pulmonary epithelial and endothelial cells.
    Article Snippet: .. 10 μg of proteins were separated in 12.5% SDS – PAGE gels and blotted onto nitrocellulose or PVDF membranes by semi-wet transfer for 10min at 2.5 A and 25 V. Membranes were blocked in TBS-T with 5% non – fat milk for 1 h in RT following incubation overnight at 4 °C with primary (1:1000) antibodies: Flag (Sigma, Cat. F1804), IL-1β (Cell Signaling Technology, MA, USA, Cat. 2022 and Affinity Biosciences, Ohio, USA, Cat. AF4012), IL-1α (Santa Cruz, CA, USA, Cat. sc-9983), NLRP3 (Adipogen, Switzerland, Cat. AG-20B-0014-C100), GSDMD (Cell Signaling Technology, Cat. 96458), β-actin (Novus, CO, USA, Cat.NB600), GAPDH (Millipore, MA, USA, Cat. MAB374; 1:5000). .. Appropriate mouse or rabbit HRP-conjugated secondary antibody (Vector Laboratories Inc., CA, USA, Cat. PI-2000, PI-1000; 1:10000) was used to detect primary antibodies.

    Incubation:

    Article Title: SARS-CoV-2 and its ORF3a, E and M viroporins activate inflammasome in human macrophages and induce of IL-1 α in pulmonary epithelial and endothelial cells
    Article Snippet: .. 10 μg of proteins were separated in 12.5% SDS – PAGE gels and blotted onto nitrocellulose or PVDF membranes by semi-wet transfer for 10 min at 2.5 A and 25 V. Membranes were blocked in TBS-T with 5% non – fat milk for 1 h in RT following incubation overnight at 4 °C with primary (1:1000) antibodies: Flag (Sigma, Cat. F1804), IL-1β (Cell Signaling Technology, MA, USA, Cat. 2022 and Affinity Biosciences, Ohio, USA, Cat. AF4012), IL-1α (Santa Cruz, CA, USA, Cat. sc-9983), NLRP3 (Adipogen, Switzerland, Cat. AG-20B-0014-C100), GSDMD (Cell Signaling Technology, Cat. 96458), β-actin (Novus, CO, USA, Cat.NB600), GAPDH (Millipore, MA, USA, Cat. MAB374; 1:5000). .. Appropriate mouse or rabbit HRP-conjugated secondary antibody (Vector Laboratories Inc., CA, USA, Cat. PI-2000, PI-1000; 1:10000) was used to detect primary antibodies.

    Article Title: SARS-CoV-2 and its ORF3a, E and M viroporins activate inflammasome in human macrophages and induce of IL-1α in pulmonary epithelial and endothelial cells.
    Article Snippet: .. 10 μg of proteins were separated in 12.5% SDS – PAGE gels and blotted onto nitrocellulose or PVDF membranes by semi-wet transfer for 10min at 2.5 A and 25 V. Membranes were blocked in TBS-T with 5% non – fat milk for 1 h in RT following incubation overnight at 4 °C with primary (1:1000) antibodies: Flag (Sigma, Cat. F1804), IL-1β (Cell Signaling Technology, MA, USA, Cat. 2022 and Affinity Biosciences, Ohio, USA, Cat. AF4012), IL-1α (Santa Cruz, CA, USA, Cat. sc-9983), NLRP3 (Adipogen, Switzerland, Cat. AG-20B-0014-C100), GSDMD (Cell Signaling Technology, Cat. 96458), β-actin (Novus, CO, USA, Cat.NB600), GAPDH (Millipore, MA, USA, Cat. MAB374; 1:5000). .. Appropriate mouse or rabbit HRP-conjugated secondary antibody (Vector Laboratories Inc., CA, USA, Cat. PI-2000, PI-1000; 1:10000) was used to detect primary antibodies.



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    93
    Santa Cruz Biotechnology mouse anti il 1α antibody
    Interleukin (IL)−1β levels are elevated in MK2/3 double-knockout (DKO) mice. ( a ) Il1b mRNA levels are increased in untreated and ( b ) <t>IL-1α-treated</t> (5 ng/mL, 1 h) MK2/3 -DKO bone marrow-derived macrophages (BMDMs) compared to wild type (WT). WT n = 14, DKO n = 13. ( c ) Basal IL-1β protein levels are elevated in MK2/3 -DKO BMDM. Elongation factor 2 (EF2) serves as a control. One representative Western blot of total WT n = 4, DKO n = 6. ( d ) The concentration of IL-1β is higher in the supernatant of untreated and ( e ) IL-1α (5 ng/mL, 4 h) + Nigericin (20 µM, 8 h) or ( f ) IL-1α + ATP (5 mM, 5.5 h)-treated MK2/3 -DKO BMDM than in WT. ( d , e ) WT n = 9, DKO n = 8. ( f ) n = 3/group. ( g ) The basal concentration of IL-1β is elevated in the serum of MK2/3 -DKO mice. n = 6 mice/group, whereby one sample/group was pooled from 3 mouse sera. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.
    Mouse Anti Il 1α Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/il+1%CE%B1/IL-1%CE%B1/pm41898739-229-78-82
    Average 93 stars, based on 1 article reviews
    mouse anti il 1α antibody - by Bioz Stars, 2026-09
    93/100 stars
    		  Buy from Supplier
    human recombinant il 1α  (R&D Systems)
    94
    R&D Systems human recombinant il 1α
    Interleukin (IL)−1β levels are elevated in MK2/3 double-knockout (DKO) mice. ( a ) Il1b mRNA levels are increased in untreated and ( b ) <t>IL-1α-treated</t> (5 ng/mL, 1 h) MK2/3 -DKO bone marrow-derived macrophages (BMDMs) compared to wild type (WT). WT n = 14, DKO n = 13. ( c ) Basal IL-1β protein levels are elevated in MK2/3 -DKO BMDM. Elongation factor 2 (EF2) serves as a control. One representative Western blot of total WT n = 4, DKO n = 6. ( d ) The concentration of IL-1β is higher in the supernatant of untreated and ( e ) IL-1α (5 ng/mL, 4 h) + Nigericin (20 µM, 8 h) or ( f ) IL-1α + ATP (5 mM, 5.5 h)-treated MK2/3 -DKO BMDM than in WT. ( d , e ) WT n = 9, DKO n = 8. ( f ) n = 3/group. ( g ) The basal concentration of IL-1β is elevated in the serum of MK2/3 -DKO mice. n = 6 mice/group, whereby one sample/group was pooled from 3 mouse sera. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.
    Human Recombinant Il 1α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/il+1%CE%B1/Recombinant+Human+IFN-alpha-1a+Protein%2C+CF/us12582628-101-3-8
    Average 94 stars, based on 1 article reviews
    human recombinant il 1α - by Bioz Stars, 2026-09
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    a, representation of the proteins with denaturing curves significantly altered (red) or unaltered (black) by 3,2-HPP treatment of THP1-Dual™ cells, using thermal proteome profiling (two combined experiments, proteomic coverage: 4301, NPARC test); b, protein-protein interaction network of proteins with denaturation curves altered by 3,2-HPP treatment (red) and the transcriptional regulators of 3, 2-HPP impacted genes in M3-9-M tumours (yellow); c, impact of 3,2-HPP treatment of THP1-Dual™ cells on the denaturation curve of GSDMD (two-combined experiments, NPARC test); d, NF-κB induction in THP1-Dual™ reporter cells treated with conditioned media harvested from WT or Gsdmd- KO THP1 cells treated with LPS ± HPP metabolites ± IL-1α and −1β neutralizing antibody (three combined experiments, one-way ANOVA with Bonferroni multiple comparison test); e-h, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT or Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); i-l, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT vs. Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS and NG ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); m, western blot of THP-1 cells treated with LPS ± NG ± HPPs (representative of two experiments).

    Journal: bioRxiv

    Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

    doi: 10.64898/2026.04.23.720410

    Figure Lengend Snippet: a, representation of the proteins with denaturing curves significantly altered (red) or unaltered (black) by 3,2-HPP treatment of THP1-Dual™ cells, using thermal proteome profiling (two combined experiments, proteomic coverage: 4301, NPARC test); b, protein-protein interaction network of proteins with denaturation curves altered by 3,2-HPP treatment (red) and the transcriptional regulators of 3, 2-HPP impacted genes in M3-9-M tumours (yellow); c, impact of 3,2-HPP treatment of THP1-Dual™ cells on the denaturation curve of GSDMD (two-combined experiments, NPARC test); d, NF-κB induction in THP1-Dual™ reporter cells treated with conditioned media harvested from WT or Gsdmd- KO THP1 cells treated with LPS ± HPP metabolites ± IL-1α and −1β neutralizing antibody (three combined experiments, one-way ANOVA with Bonferroni multiple comparison test); e-h, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT or Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); i-l, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT vs. Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS and NG ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); m, western blot of THP-1 cells treated with LPS ± NG ± HPPs (representative of two experiments).

    Article Snippet: To neutralize IL-1 receptor signalling, the following antibodies were used: anti-human IL-1α (1 μg/mL; clone 7D4; mabg-hil1a-3; InvivoGen), anti-human IL-1β (1 μg/mL; clone 4H5; mabg-hil1b-3; InvivoGen) and IgG1 isotype control (1 μg/mL; clone T8E5; mabg1-ctrlm; InvivoGen).

    Techniques: Comparison, Cell Culture, Western Blot

    Interleukin (IL)−1β levels are elevated in MK2/3 double-knockout (DKO) mice. ( a ) Il1b mRNA levels are increased in untreated and ( b ) IL-1α-treated (5 ng/mL, 1 h) MK2/3 -DKO bone marrow-derived macrophages (BMDMs) compared to wild type (WT). WT n = 14, DKO n = 13. ( c ) Basal IL-1β protein levels are elevated in MK2/3 -DKO BMDM. Elongation factor 2 (EF2) serves as a control. One representative Western blot of total WT n = 4, DKO n = 6. ( d ) The concentration of IL-1β is higher in the supernatant of untreated and ( e ) IL-1α (5 ng/mL, 4 h) + Nigericin (20 µM, 8 h) or ( f ) IL-1α + ATP (5 mM, 5.5 h)-treated MK2/3 -DKO BMDM than in WT. ( d , e ) WT n = 9, DKO n = 8. ( f ) n = 3/group. ( g ) The basal concentration of IL-1β is elevated in the serum of MK2/3 -DKO mice. n = 6 mice/group, whereby one sample/group was pooled from 3 mouse sera. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

    doi: 10.3390/ijms27073232

    Figure Lengend Snippet: Interleukin (IL)−1β levels are elevated in MK2/3 double-knockout (DKO) mice. ( a ) Il1b mRNA levels are increased in untreated and ( b ) IL-1α-treated (5 ng/mL, 1 h) MK2/3 -DKO bone marrow-derived macrophages (BMDMs) compared to wild type (WT). WT n = 14, DKO n = 13. ( c ) Basal IL-1β protein levels are elevated in MK2/3 -DKO BMDM. Elongation factor 2 (EF2) serves as a control. One representative Western blot of total WT n = 4, DKO n = 6. ( d ) The concentration of IL-1β is higher in the supernatant of untreated and ( e ) IL-1α (5 ng/mL, 4 h) + Nigericin (20 µM, 8 h) or ( f ) IL-1α + ATP (5 mM, 5.5 h)-treated MK2/3 -DKO BMDM than in WT. ( d , e ) WT n = 9, DKO n = 8. ( f ) n = 3/group. ( g ) The basal concentration of IL-1β is elevated in the serum of MK2/3 -DKO mice. n = 6 mice/group, whereby one sample/group was pooled from 3 mouse sera. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

    Techniques: Double Knockout, Derivative Assay, Control, Western Blot, Concentration Assay

    The level of IL-1β is increased in immortalized MK2 -KO (i MK2 -KO) cells. ( a ) i MK2 -KO cells transduced with an empty vector (+ GFP ) showed elevated levels of Il1b mRNA compared to MK2 -rescued cells (+ MK2 ) (right, n = 8), similar to those observed in MK2/3 -DKO and WT bone-marrow-derived macrophages (BMDMs) (left; WT n = 9, DKO n = 8). ( b ) i MK2 -KO + GFP cells show increased Il1b mRNA after IL-1α treatment (5 ng/mL) compared to MK2 -rescued cells. n = 3. ( c ) RAW cells treated with MK2 siRNA have higher Il1b mRNA levels compared to the control after IL-1α stimulation. ( d ) Similar to MK2 , rescuing MK3 decreases the level of Il1b mRNA in resting or ( e ) IL-1α (5 ng/mL, 1 h)-treated i MK2 -KO cells, ( f ) as well as the basal level of IL-1β protein. Histone H3 serves as a control. ( a , c ) Student’s t -test, ( b ) 2W-RM-ANOVA with Bonferroni posttests, ( d , e ) 1W-ANOVA with Tukey’s Multiple Comparison Test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

    doi: 10.3390/ijms27073232

    Figure Lengend Snippet: The level of IL-1β is increased in immortalized MK2 -KO (i MK2 -KO) cells. ( a ) i MK2 -KO cells transduced with an empty vector (+ GFP ) showed elevated levels of Il1b mRNA compared to MK2 -rescued cells (+ MK2 ) (right, n = 8), similar to those observed in MK2/3 -DKO and WT bone-marrow-derived macrophages (BMDMs) (left; WT n = 9, DKO n = 8). ( b ) i MK2 -KO + GFP cells show increased Il1b mRNA after IL-1α treatment (5 ng/mL) compared to MK2 -rescued cells. n = 3. ( c ) RAW cells treated with MK2 siRNA have higher Il1b mRNA levels compared to the control after IL-1α stimulation. ( d ) Similar to MK2 , rescuing MK3 decreases the level of Il1b mRNA in resting or ( e ) IL-1α (5 ng/mL, 1 h)-treated i MK2 -KO cells, ( f ) as well as the basal level of IL-1β protein. Histone H3 serves as a control. ( a , c ) Student’s t -test, ( b ) 2W-RM-ANOVA with Bonferroni posttests, ( d , e ) 1W-ANOVA with Tukey’s Multiple Comparison Test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

    Techniques: Transduction, Plasmid Preparation, Derivative Assay, Control, Comparison

    The non-canonical NF-κB pathway is activated in i MK2 -KO cells (mRNA). ( a ) Inhibition of the canonical NF-κB pathway using the IKKβ inhibitors Takinib (10 µM, 2 h) and sc-514 (10 µM, 2 h) reduced Il1b mRNA levels in IL-1α- and LPS-treated i MK2 -KO cells, but did not affect basal Il1b levels. Inhibiting IKKα and IKKβ with HPN-01 (10 µM, 2 h) reduced Il1b mRNA levels in untreated (UT) and IL-1α (5 ng/mL, 1 h)- or LPS-stimulated cells (100 ng/mL, 1 h). Inhibition of the non-canonical NF-κB pathway by IKKα inhibitor B022 (5 µM, 2 h) mainly reduced basal Il1b mRNA. ( b , c ) IL-1β protein level is reduced in resting i MK2 -KO cells after treatment with B022 (5 µM, 7 h). GAPDH serves as a control. ( d ) Il1b mRNA is reduced in MK2/3 -DKO BMDMs after treatment with B022 (5 µM, 2 h). ( e ) The level of Map3k14 mRNA is increased in i MK2 -KO + GFP . ( f ) Relb mRNA level is increased in UT i MK2 -KO + GFP cells. ( g ) Relb and ( h ) Nfkb2 mRNA levels are increased in IL-1α (5 ng/mL, 1 h)-stimulated i MK2 -KO + GFP cells. ( i ) Basal Traf2 mRNA is reduced in i MK2 -KO + GFP cells. ( j ) The Traf3 mRNA level is not changed significantly. ( a ) 1W-ANOVA with Tukey’s Multiple Comparison Test, ( c – j ) Student’s t -test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

    doi: 10.3390/ijms27073232

    Figure Lengend Snippet: The non-canonical NF-κB pathway is activated in i MK2 -KO cells (mRNA). ( a ) Inhibition of the canonical NF-κB pathway using the IKKβ inhibitors Takinib (10 µM, 2 h) and sc-514 (10 µM, 2 h) reduced Il1b mRNA levels in IL-1α- and LPS-treated i MK2 -KO cells, but did not affect basal Il1b levels. Inhibiting IKKα and IKKβ with HPN-01 (10 µM, 2 h) reduced Il1b mRNA levels in untreated (UT) and IL-1α (5 ng/mL, 1 h)- or LPS-stimulated cells (100 ng/mL, 1 h). Inhibition of the non-canonical NF-κB pathway by IKKα inhibitor B022 (5 µM, 2 h) mainly reduced basal Il1b mRNA. ( b , c ) IL-1β protein level is reduced in resting i MK2 -KO cells after treatment with B022 (5 µM, 7 h). GAPDH serves as a control. ( d ) Il1b mRNA is reduced in MK2/3 -DKO BMDMs after treatment with B022 (5 µM, 2 h). ( e ) The level of Map3k14 mRNA is increased in i MK2 -KO + GFP . ( f ) Relb mRNA level is increased in UT i MK2 -KO + GFP cells. ( g ) Relb and ( h ) Nfkb2 mRNA levels are increased in IL-1α (5 ng/mL, 1 h)-stimulated i MK2 -KO + GFP cells. ( i ) Basal Traf2 mRNA is reduced in i MK2 -KO + GFP cells. ( j ) The Traf3 mRNA level is not changed significantly. ( a ) 1W-ANOVA with Tukey’s Multiple Comparison Test, ( c – j ) Student’s t -test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

    Techniques: Inhibition, Control, Comparison

    The non-canonical NF-κB pathway is activated in i MK2 -KO cells (protein). ( a – c ) Compared to i MK2 - KO + MK2 cells, untreated (UT) i MK2 -KO + GFP cells have higher levels of the non-canonical proteins RelB and NF-κB2 in nuclear and cytoplasmic fractions, but not of the canonical RelA and NF-κB1 proteins. Following IL-1α treatment (5 ng/mL, 2 h), the nuclear fraction of i MK2 -KO cells showed elevated protein levels of RelB, NF-κB2, RelA, and NF-κB1. p53 and GAPDH serve as controls for successful nuclear/cytoplasmic separation. EF2 acts as a general control, used for normalization. ( d , e ) The basal protein level of TRAF2 is reduced in whole-cell lysis of i MK2 -KO + GFP cells, whereas TRAF3 and cIAP1/2 are not changed. ( f ) The protein level of c-Rel is increased in i MK2 -KO+ GFP cells. Examples of Western blots from different gels are shown. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

    doi: 10.3390/ijms27073232

    Figure Lengend Snippet: The non-canonical NF-κB pathway is activated in i MK2 -KO cells (protein). ( a – c ) Compared to i MK2 - KO + MK2 cells, untreated (UT) i MK2 -KO + GFP cells have higher levels of the non-canonical proteins RelB and NF-κB2 in nuclear and cytoplasmic fractions, but not of the canonical RelA and NF-κB1 proteins. Following IL-1α treatment (5 ng/mL, 2 h), the nuclear fraction of i MK2 -KO cells showed elevated protein levels of RelB, NF-κB2, RelA, and NF-κB1. p53 and GAPDH serve as controls for successful nuclear/cytoplasmic separation. EF2 acts as a general control, used for normalization. ( d , e ) The basal protein level of TRAF2 is reduced in whole-cell lysis of i MK2 -KO + GFP cells, whereas TRAF3 and cIAP1/2 are not changed. ( f ) The protein level of c-Rel is increased in i MK2 -KO+ GFP cells. Examples of Western blots from different gels are shown. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

    Techniques: Control, Lysis, Western Blot

    The non-canonical NF-κB pathway is activated in i MK2 -KO cells. RNA sequencing revealed increased levels of mRNA for components of the IL-1β and non-canonical NF-κB pathways, as well as for targets of the non-canonical NF-κB pathway, in i MK2 -KO + GFP cells. This finding was reinforced by IL-1α treatment (5 ng/mL, 1 h).

    Journal: International Journal of Molecular Sciences

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

    doi: 10.3390/ijms27073232

    Figure Lengend Snippet: The non-canonical NF-κB pathway is activated in i MK2 -KO cells. RNA sequencing revealed increased levels of mRNA for components of the IL-1β and non-canonical NF-κB pathways, as well as for targets of the non-canonical NF-κB pathway, in i MK2 -KO + GFP cells. This finding was reinforced by IL-1α treatment (5 ng/mL, 1 h).

    Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

    Techniques: RNA Sequencing

    The MK2 kinase activity is not involved, but the MK2 C-terminus is important. ( a ) The rescued MK2 kinase-inactive mutant, MK2K79R, reduces Il1b mRNA levels to a degree comparable to that of the rescued MK2 in untreated (UT) and ( b ) IL-1α-treated (5 ng/mL, 1 h) i MK2 -KO macrophages. ( c ) i MK2 -KO cells have lower levels of the p38α protein. These levels can be restored by rescuing MK2 , but not by rescuing a MK2 mutant lacking the C-terminus MK2-Δ365–386 . ( d ) MK2-Δ365-386 does not affect the Il1b mRNA levels in IL-1α-treated cells. 1W-ANOVA with Tukey’s Multiple Comparison Test, mean ± SEM, ** p < 0.01, *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

    doi: 10.3390/ijms27073232

    Figure Lengend Snippet: The MK2 kinase activity is not involved, but the MK2 C-terminus is important. ( a ) The rescued MK2 kinase-inactive mutant, MK2K79R, reduces Il1b mRNA levels to a degree comparable to that of the rescued MK2 in untreated (UT) and ( b ) IL-1α-treated (5 ng/mL, 1 h) i MK2 -KO macrophages. ( c ) i MK2 -KO cells have lower levels of the p38α protein. These levels can be restored by rescuing MK2 , but not by rescuing a MK2 mutant lacking the C-terminus MK2-Δ365–386 . ( d ) MK2-Δ365-386 does not affect the Il1b mRNA levels in IL-1α-treated cells. 1W-ANOVA with Tukey’s Multiple Comparison Test, mean ± SEM, ** p < 0.01, *** p < 0.001.

    Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

    Techniques: Activity Assay, Mutagenesis, Comparison

    p38α inactivates the non-canonical NF-κB pathway independent of the kinase activity. ( a – c ) Overexpression of p38α in i MK2 -KO cells increases basal TRAF2 and reduces basal RelB protein levels. ( d ) Overexpression of p38α and kinase inactive mutant p38-AGF reduce basal Il1b and ( e ) Map3k14 mRNA and ( f ) increase basal Traf2 mRNA in resting i MK2 -KO cells. ( g ) Relb and ( h ) Nfkb2 mRNA are reduced in IL-1α-treated (5 ng/mL, 1 h) i MK2 -KO+ p38α and +p38-AGF cells. ( b , c ) Student’s t -test, ( d – h ) 1W-ANOVA followed by Tukey’s Multiple Comparison Test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

    doi: 10.3390/ijms27073232

    Figure Lengend Snippet: p38α inactivates the non-canonical NF-κB pathway independent of the kinase activity. ( a – c ) Overexpression of p38α in i MK2 -KO cells increases basal TRAF2 and reduces basal RelB protein levels. ( d ) Overexpression of p38α and kinase inactive mutant p38-AGF reduce basal Il1b and ( e ) Map3k14 mRNA and ( f ) increase basal Traf2 mRNA in resting i MK2 -KO cells. ( g ) Relb and ( h ) Nfkb2 mRNA are reduced in IL-1α-treated (5 ng/mL, 1 h) i MK2 -KO+ p38α and +p38-AGF cells. ( b , c ) Student’s t -test, ( d – h ) 1W-ANOVA followed by Tukey’s Multiple Comparison Test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

    Techniques: Activity Assay, Over Expression, Mutagenesis, Comparison