il-1α Search Results


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  • 93
    Millipore interleukin 1α il 1α
    Effect of S -nitroso- N -acetyl-penicillamine (SNAP) with <t>interleukin-1α</t> (IL-1α) on the production of nitric oxide (NO) in trabecular meshwork (TM) cells. SNAP significantly increased NO production in a dose-dependent manner ( * p
    Interleukin 1α Il 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems interleukin 1α il 1α
    Secreted levels of hBD-2, <t>IL-1α</t> and IL-8 in the unstimulated GECs of various genotypes are determined by ELISA. Each experiment was performed three times, in duplicate. The data are presented as mean+SD. *p
    Interleukin 1α Il 1α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 88/100, based on 247 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    tebu-bio sa cytokine interleukin 1α il 1α
    Hybrid IGKV sequences from multiplex PCR IGKV - IGKJ 5. Examples of sequences from the cloned PCR products obtained from <t>interleukin-1α</t> (IL-1α) or lipopolysaccharide (LPS) -treated human pre-B 697 cells. Seven chimeric sequences were selected
    Cytokine Interleukin 1α Il 1α, supplied by tebu-bio sa, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam interleukin 1α il 1α
    Hybrid IGKV sequences from multiplex PCR IGKV - IGKJ 5. Examples of sequences from the cloned PCR products obtained from <t>interleukin-1α</t> (IL-1α) or lipopolysaccharide (LPS) -treated human pre-B 697 cells. Seven chimeric sequences were selected
    Interleukin 1α Il 1α, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genzyme mouse interleukin 1α il 1α
    Hybrid IGKV sequences from multiplex PCR IGKV - IGKJ 5. Examples of sequences from the cloned PCR products obtained from <t>interleukin-1α</t> (IL-1α) or lipopolysaccharide (LPS) -treated human pre-B 697 cells. Seven chimeric sequences were selected
    Mouse Interleukin 1α Il 1α, supplied by Genzyme, used in various techniques. Bioz Stars score: 80/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    PromoCell il 1α
    Neutralization of IFN-β and <t>IL-1α</t> in conditioned medium from C. pneumoniae -infected WISH cells. Conditioned medium was incubated with neutralizing antibodies against IFN-β (A) or IL-1α (B) and then used to study the effect
    Il 1α, supplied by PromoCell, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology il 1α
    Experimental evidence that regulation of parkin expression and phosphorylation is related to glial activation and excess IL-1β in Alzheimer brain and in AD Model Systems. a Parkin in neurons in brains of age-matched control patients (AMC) is diffusely distributed within the cytoplasm whereas parkin in neurons in brains of Alzheimer patients (AD) is present in rosette-like aggregates ( b ). c Parkin in Alzheimer brain is colocalized with tau aggregates. d Activated glia overexpressing <t>IL-1α</t> (brown) are immediately adjacent to parkin-immunoreactive neurons (blue), scale bar 20 μM. e Comparative levels of IL-1α, IL-1β, and parkin mRNAs in wild-type littermates or PD-APP mice. f Representative western immunoblot showing proteins from NT2 cells either treated, or not, overnight with IL-1β. g Phospho-parkin levels (P-parkin Ser 65 ) in two representative western immunoblots of proteins from NT2 cell and rat primary neuron cultures treated, or not, with IL-1β. h Quantification of parkin in untreated and treated cell cultures ( n = 6), data is mean ± SEM (** p
    Il 1α, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche il 1α
    Effect of TGF-β on dexamethasone (Dex, 10 nM)-induced increases in (A) IκBα, (B) SCNN1A and (C) GILZ mRNA levels after 4 h incubation with <t>IL-1α</t> (1 ng·mL −1 ). Data are presented as the mean and SEM of n = 3 experiments (each determined in triplicate) and the gene expression is normalized to concurrently measured 18 s RNA levels and referenced to levels in control/IL-1α treated cells (1.0). * P
    Il 1α, supplied by Roche, used in various techniques. Bioz Stars score: 90/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genzyme il 1α
    Characteristics of endogenous intracellular ITEGE neoepitope generated by bovine articular chondrocytes. Bovine articular chondrocytes were released from alginate beads following a 48-hour pretreatment with 10 ng/ml <t>interleukin-1α.</t> A and B , Overlay
    Il 1α, supplied by Genzyme, used in various techniques. Bioz Stars score: 92/100, based on 102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher il 1α
    CM from <t>IL-1α</t> deficient cells fails to support cancer cell invasiveness. The Boyden chamber invasion assay was used to determine the effects of CM from senescent HCA2 cells infected with shGFP or shIL-1α-expressing (#1 and #3) lentiviruses
    Il 1α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 877 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam il 1α
    TRAIL expression and modulation in pIL6-TRAIL + -GFP + -UC-MSCs. a Left: transfection efficiency of UC-MSCs evaluated by flow cytometry for GFP fluorescence. Wildtype UC-MSCs were negative as compared to GFP + -UC-MSCs (red histogram: 83.5%) in a similar amount as for pIL6-TRAIL + -GFP + -UC-MSCs (blue histogram: 87.6%). Right: FACS analysis revealed constitutive TRAIL expression in transduced UC-MSCs (green histogram: 62.5%) that was significantly enhanced by the conditioned medium from U-266 cells (blue histogram: 80.3%) as well as in response to <t>IL-1α/IL-1β</t> (red histogram: 91.2%). b Confocal microscopy of pIL6-TRAIL + -GFP + -UC-MSCs confirmed the upregulation of TRAIL (FITC, green) induced by either U-266 supernatant or IL-1α/IL-1β treatment. Phalloidin staining was used to show actin (TRITC-red); nuclei were counterstained with Hoechst 33342 (blue). Magnification: 100×. c Evaluation of TRAIL expression in transduced cells by RT-qPCR, western blot analysis, and ELISA. (i) After either U-266 or IL-1α/IL-1β treatment, TRAIL mRNA levels in pIL6-TRAIL + -GFP + -UC-MSCs increased up to 2.2-fold and 2.5-fold compared to basal condition. Data are relative amount of mRNA expression normalized to GAPDH and are presented as mean ± SD of three independent experiments. * p
    Il 1α, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc il 1α
    Melatonin inhibits p65 phosphorylation and nuclear translocation in thyroid carcinoma. (A) Western blot analysis of nuclear NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells treated with melatonin for 24 h. (B) ELISA analysis of NF-κB/p65 activity in 8505c, TPC-1 and KTC-1 cells after treatment with melatonin for 24 h. (C) Expression of NF-κB/p65 response genes in 8505c, TPC-1 and KTC-1 cells was detected by qPCR assays. (D) Immunoblotting of <t>IL-1α,</t> Cyclin D1 and MMP9 in 8505c, TPC-1 and KTC-1 cells after treatment with melatonin (2 mM) for 24 h. β-Actin was used as a loading control. (E) Immunoblotting of NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells treated with or without TNFα (10 ng/mL) and melatonin (2 mM) for 24 h. β-Actin was used as a loading control. * P
    Il 1α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 1α
    Spa spring water impairs pro-inflammatory cytokine production on HaCaT cells under TLR stimulation. The HaCaT cells were treated with TLR1 through TLR6 agonist for 1, 4, 10, and 24 hours, respectively. Spa spring water diluted 1 : 30 fold was added prior to adding TLR agonists or simultaneously. After the indicated time, the supernatants were collected to measure secreted level of IL-6 (A), IL-8 (B), TNF-α (C), <t>IL-1α</t> (D), and GM-CSF (E) via ELISA. The data are expressed as the means±SEM ( * p
    Il 1α, supplied by BioLegend, used in various techniques. Bioz Stars score: 93/100, based on 196 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PBL Assay il 1α
    Spa spring water impairs pro-inflammatory cytokine production on HaCaT cells under TLR stimulation. The HaCaT cells were treated with TLR1 through TLR6 agonist for 1, 4, 10, and 24 hours, respectively. Spa spring water diluted 1 : 30 fold was added prior to adding TLR agonists or simultaneously. After the indicated time, the supernatants were collected to measure secreted level of IL-6 (A), IL-8 (B), TNF-α (C), <t>IL-1α</t> (D), and GM-CSF (E) via ELISA. The data are expressed as the means±SEM ( * p
    Il 1α, supplied by PBL Assay, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Miltenyi Biotec il 1α
    Spa spring water impairs pro-inflammatory cytokine production on HaCaT cells under TLR stimulation. The HaCaT cells were treated with TLR1 through TLR6 agonist for 1, 4, 10, and 24 hours, respectively. Spa spring water diluted 1 : 30 fold was added prior to adding TLR agonists or simultaneously. After the indicated time, the supernatants were collected to measure secreted level of IL-6 (A), IL-8 (B), TNF-α (C), <t>IL-1α</t> (D), and GM-CSF (E) via ELISA. The data are expressed as the means±SEM ( * p
    Il 1α, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biogenesis Inc il 1α
    Spa spring water impairs pro-inflammatory cytokine production on HaCaT cells under TLR stimulation. The HaCaT cells were treated with TLR1 through TLR6 agonist for 1, 4, 10, and 24 hours, respectively. Spa spring water diluted 1 : 30 fold was added prior to adding TLR agonists or simultaneously. After the indicated time, the supernatants were collected to measure secreted level of IL-6 (A), IL-8 (B), TNF-α (C), <t>IL-1α</t> (D), and GM-CSF (E) via ELISA. The data are expressed as the means±SEM ( * p
    Il 1α, supplied by Biogenesis Inc, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery il 1α
    <t>IL-1α</t> signals through IL-1R to induce the SASP. (A) Western blot analysis of IL-1α, IL-1β, and tubulin (Tub) levels in IMR90T cells expressing either a doxycycline-inducible scramble shRNA (shScr) or shRNA against IL-1α (shIL1α). Cells were treated with either ethanol (Et) as a control or tamoxifen (4OHT) to induce Ras activity and doxycycline to induce knockdown. (B) qRT-PCR analyses of the indicated genes in the indicated cells. Values are shown as fold change compared to levels in shScr treated with Et. (C) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (D) Western blot analysis detecting IL-1α and Tub levels in wild-type (WT) and IL-1α-deleted (KO) IMR90T cells. WT1, parental cell line; WT2, a clone that failed to delete IL-1α; KO pool, total pool of clones, where KO1 and KO2 each represent an independent clone. (E) qRT-PCR detecting IL-8 mRNA levels in the indicated cells. (F) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (G) qRT-PCR analyses of the indicated SASP factors found via transcriptome analysis to be affected by IL-1R knockdown. Data are represented as fold change compared to levels for WT1 treated with Et. (H) Western blot analysis of IL-1α in IMR90T cells expressing an empty vector control (EV), full-length IL-1α (IL-1α FL), the N-terminal propiece (ppIL-1α), or the C-terminal mature fragment (mIL-1α). The antibody used only recognizes the C-terminal portion of IL-1α. (I) qRT-PCR analyses of the indicated genes in the indicated cell lines. Primers for IL-1α anneal to the N terminus. Values are shown as fold change compared to levels in cells expressing EV. Each experiment was performed at least 3 times. ns, not significant; *, P
    Il 1α, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Immunotec il 1α
    <t>IL-1α</t> signals through IL-1R to induce the SASP. (A) Western blot analysis of IL-1α, IL-1β, and tubulin (Tub) levels in IMR90T cells expressing either a doxycycline-inducible scramble shRNA (shScr) or shRNA against IL-1α (shIL1α). Cells were treated with either ethanol (Et) as a control or tamoxifen (4OHT) to induce Ras activity and doxycycline to induce knockdown. (B) qRT-PCR analyses of the indicated genes in the indicated cells. Values are shown as fold change compared to levels in shScr treated with Et. (C) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (D) Western blot analysis detecting IL-1α and Tub levels in wild-type (WT) and IL-1α-deleted (KO) IMR90T cells. WT1, parental cell line; WT2, a clone that failed to delete IL-1α; KO pool, total pool of clones, where KO1 and KO2 each represent an independent clone. (E) qRT-PCR detecting IL-8 mRNA levels in the indicated cells. (F) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (G) qRT-PCR analyses of the indicated SASP factors found via transcriptome analysis to be affected by IL-1R knockdown. Data are represented as fold change compared to levels for WT1 treated with Et. (H) Western blot analysis of IL-1α in IMR90T cells expressing an empty vector control (EV), full-length IL-1α (IL-1α FL), the N-terminal propiece (ppIL-1α), or the C-terminal mature fragment (mIL-1α). The antibody used only recognizes the C-terminal portion of IL-1α. (I) qRT-PCR analyses of the indicated genes in the indicated cell lines. Primers for IL-1α anneal to the N terminus. Values are shown as fold change compared to levels in cells expressing EV. Each experiment was performed at least 3 times. ns, not significant; *, P
    Il 1α, supplied by Immunotec, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson il 1α
    CD11a hi FcγRIII hi B cells produce a high level of IFN-γ in response to CD40 ligation. C57BL/6 mice were infected with 2 × 10 6 LM. (A) The intracellular expression of <t>IL-1α,</t> IL-2, IL-6, IL-10 and IFN-γ in CD11a hi FcγRIII hi and conventional B cells was assayed on day 3 post-infection. (B - G) Splenic CD11a hi FcγRIII hi and conventional B cells were sorted on day 3 post-infection. IL-1β, IL-2, IL-6, IL-12p70, IFN-γ, or TNF-α secretion by CD11a hi FcγRIII hi and conventional B cells was detected by ELISA. Data shown represent the mean ± SD of triplicate experiments. * P
    Il 1α, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 403 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    FUJIFILM il 1α
    Effect of <t>IL-1α,</t> IL-1α antibody and IL-1R1 antagonist on the expression of IL-1R1 and S100A9 mRNA in HaCaT cells
    Il 1α, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson paired interleukin 1α il 1α antibodies
    Effect of <t>IL-1α,</t> IL-1α antibody and IL-1R1 antagonist on the expression of IL-1R1 and S100A9 mRNA in HaCaT cells
    Paired Interleukin 1α Il 1α Antibodies, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Epitomics il 1α
    Effect of <t>IL-1α,</t> IL-1α antibody and IL-1R1 antagonist on the expression of IL-1R1 and S100A9 mRNA in HaCaT cells
    Il 1α, supplied by Epitomics, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech il 1α
    Secreted MMP activity of STEs and cartilage explants cultured alone or together. After 7 days, the MMP-activity (rfu/sec) was determined (in culture medium) of normal ( A, B ) and OA ( C, D ) STEs and healthy cartilage explants, cultured alone or together, without ( A, C ) or with ( B, D ) <t>IL-1α</t> stimulation. Each line represents an individual donor and demonstrates the activity of STEs alone (left), STEs together with cartilage explants (middle) or cartilage explants cultured alone (right). The MMP activity in the medium of STEs cultured alone was higher, compared to the MMP activity in the co-culture conditions. Significance levels are indicated.
    Il 1α, supplied by PeproTech, used in various techniques. Bioz Stars score: 94/100, based on 324 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore il 1α
    <t>IL-1α</t> localization and knockdown in THP-1 macrophages. A ) THP-1 macrophages were transfected with control (ctr) or IL-1α siRNA for 48 h, followed by 24 h stimulation with MSU (100 µg/ml). Cell lysates were used to examine the expression of pro-IL-1α, pro-IL-1β, cleaved capase-1, and NLRP3 in IL-1α knockdown cells. B ) THP-1 macrophages were stimulated with MSU (100 μg/ml) for 30 min. Nuclear and cyotsolic fractions were prepared to evaluate the expression of pro–IL-1α and –IL-1β. C ) Conditioned medium was collected and used for the quantification of IL-1β, TNF-α, IL-8, and MCP-1 production with without MSU stimulation. NS, nonstimulated. * P
    Il 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 420 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of S -nitroso- N -acetyl-penicillamine (SNAP) with interleukin-1α (IL-1α) on the production of nitric oxide (NO) in trabecular meshwork (TM) cells. SNAP significantly increased NO production in a dose-dependent manner ( * p

    Journal: Korean Journal of Ophthalmology : KJO

    Article Title: Effect of Nitric Oxide on the Expression of Matrix Metalloproteinase and Its Association with Migration of Cultured Trabecular Meshwork Cells

    doi: 10.3341/kjo.2016.30.1.66

    Figure Lengend Snippet: Effect of S -nitroso- N -acetyl-penicillamine (SNAP) with interleukin-1α (IL-1α) on the production of nitric oxide (NO) in trabecular meshwork (TM) cells. SNAP significantly increased NO production in a dose-dependent manner ( * p

    Article Snippet: Materials S -nitroso-N -acetyl-penicillamine (SNAP), 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT),N ω -nitro-L-arginine methyl ester (L-NAME), Griess reagent, and interleukin-1α(IL-1α) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques:

    Effect of N ω -nitro-L-arginine methyl ester (L-NAME), interleukin-1α (IL-1α), and S -nitroso- N -acetyl-penicillamine (SNAP) on the survival of trabecular meshwork cells. L-NAME, IL-1α, and SNAP did not have different effects on survival compared with the control ( p > 0.05). Values are presented as means ± standard error of the mean.

    Journal: Korean Journal of Ophthalmology : KJO

    Article Title: Effect of Nitric Oxide on the Expression of Matrix Metalloproteinase and Its Association with Migration of Cultured Trabecular Meshwork Cells

    doi: 10.3341/kjo.2016.30.1.66

    Figure Lengend Snippet: Effect of N ω -nitro-L-arginine methyl ester (L-NAME), interleukin-1α (IL-1α), and S -nitroso- N -acetyl-penicillamine (SNAP) on the survival of trabecular meshwork cells. L-NAME, IL-1α, and SNAP did not have different effects on survival compared with the control ( p > 0.05). Values are presented as means ± standard error of the mean.

    Article Snippet: Materials S -nitroso-N -acetyl-penicillamine (SNAP), 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT),N ω -nitro-L-arginine methyl ester (L-NAME), Griess reagent, and interleukin-1α(IL-1α) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques:

    Secreted levels of hBD-2, IL-1α and IL-8 in the unstimulated GECs of various genotypes are determined by ELISA. Each experiment was performed three times, in duplicate. The data are presented as mean+SD. *p

    Journal: Molecular immunology

    Article Title: Genotype-Specific Regulation of Oral Innate Immunity by T2R38 Taste Receptor

    doi: 10.1016/j.molimm.2015.10.012

    Figure Lengend Snippet: Secreted levels of hBD-2, IL-1α and IL-8 in the unstimulated GECs of various genotypes are determined by ELISA. Each experiment was performed three times, in duplicate. The data are presented as mean+SD. *p

    Article Snippet: Resultant cell culture supernatants were collected and corresponding ELISA kits were used to measure the secretion levels of IL-8, IL-1α (R & D Systems, Minneapolis, MN) and hBD-2 (PeproTech, Rocky Hill, NJ).

    Techniques: Enzyme-linked Immunosorbent Assay

    ELISA data show the changes in secreted levels of IL-1α and IL-8 in response to S. mutans when T2R38 is silenced with siRNA (T2RSi). The figure suggests that the role of T2R38 in cytokine secretion is genotype-dependent. GECs transfected with control non-silencing (NS) siRNA and subsequently stimulated with S. mutans and unstimulated GECs served as controls. Each experiment was performed three times, in duplicate. The data are presented as mean+SD. Each asterisk denotes significant difference compared with NS siRNA plus bacteria controls (p

    Journal: Molecular immunology

    Article Title: Genotype-Specific Regulation of Oral Innate Immunity by T2R38 Taste Receptor

    doi: 10.1016/j.molimm.2015.10.012

    Figure Lengend Snippet: ELISA data show the changes in secreted levels of IL-1α and IL-8 in response to S. mutans when T2R38 is silenced with siRNA (T2RSi). The figure suggests that the role of T2R38 in cytokine secretion is genotype-dependent. GECs transfected with control non-silencing (NS) siRNA and subsequently stimulated with S. mutans and unstimulated GECs served as controls. Each experiment was performed three times, in duplicate. The data are presented as mean+SD. Each asterisk denotes significant difference compared with NS siRNA plus bacteria controls (p

    Article Snippet: Resultant cell culture supernatants were collected and corresponding ELISA kits were used to measure the secretion levels of IL-8, IL-1α (R & D Systems, Minneapolis, MN) and hBD-2 (PeproTech, Rocky Hill, NJ).

    Techniques: Enzyme-linked Immunosorbent Assay, Transfection

    IL-1α and IL-8 ELISA show that the role of T2R38 in cytokine secretion is bacteria-specific and genotype-dependent. GECs transfected with control non-silencing (NS) siRNA and subsequently stimulated with different bacteria and unstimulated GECs served as controls. Each experiment was performed three times, in duplicate. The data are presented as mean+SD. Each asterisk denotes significant difference compared with NS siRNA plus bacteria controls (p

    Journal: Molecular immunology

    Article Title: Genotype-Specific Regulation of Oral Innate Immunity by T2R38 Taste Receptor

    doi: 10.1016/j.molimm.2015.10.012

    Figure Lengend Snippet: IL-1α and IL-8 ELISA show that the role of T2R38 in cytokine secretion is bacteria-specific and genotype-dependent. GECs transfected with control non-silencing (NS) siRNA and subsequently stimulated with different bacteria and unstimulated GECs served as controls. Each experiment was performed three times, in duplicate. The data are presented as mean+SD. Each asterisk denotes significant difference compared with NS siRNA plus bacteria controls (p

    Article Snippet: Resultant cell culture supernatants were collected and corresponding ELISA kits were used to measure the secretion levels of IL-8, IL-1α (R & D Systems, Minneapolis, MN) and hBD-2 (PeproTech, Rocky Hill, NJ).

    Techniques: Enzyme-linked Immunosorbent Assay, Transfection

    Hybrid IGKV sequences from multiplex PCR IGKV - IGKJ 5. Examples of sequences from the cloned PCR products obtained from interleukin-1α (IL-1α) or lipopolysaccharide (LPS) -treated human pre-B 697 cells. Seven chimeric sequences were selected

    Journal: Immunology

    Article Title: Characterization of a new V gene replacement in the absence of activation-induced cytidine deaminase and its contribution to human B-cell receptor diversity

    doi: 10.1111/imm.12192

    Figure Lengend Snippet: Hybrid IGKV sequences from multiplex PCR IGKV - IGKJ 5. Examples of sequences from the cloned PCR products obtained from interleukin-1α (IL-1α) or lipopolysaccharide (LPS) -treated human pre-B 697 cells. Seven chimeric sequences were selected

    Article Snippet: After overnight serum deprivation, 697 cells cultured at a density of 0·5 × 106 cells/ml were induced by incubation for 7 days with 10 ng/ml of the cytokine interleukin-1α (IL-1α) (Tebu-Bio, Le Perray-en-Yvelines, France) or with 1 μg/ml lipopolysaccharide (LPS, Sigma, St Louis, MO).

    Techniques: Multiplex Assay, Polymerase Chain Reaction, Clone Assay

    Neutralization of IFN-β and IL-1α in conditioned medium from C. pneumoniae -infected WISH cells. Conditioned medium was incubated with neutralizing antibodies against IFN-β (A) or IL-1α (B) and then used to study the effect

    Journal:

    Article Title: Host Cell Cytokines Induced by Chlamydia pneumoniae Decrease the Expression of Interstitial Collagens and Fibronectin in Fibroblasts ▿

    doi: 10.1128/IAI.00566-08

    Figure Lengend Snippet: Neutralization of IFN-β and IL-1α in conditioned medium from C. pneumoniae -infected WISH cells. Conditioned medium was incubated with neutralizing antibodies against IFN-β (A) or IL-1α (B) and then used to study the effect

    Article Snippet: Fibroblasts grown in six-well plates were stimulated with recombinant IFN-β (PromoCell) or IL-1α (PromoCell) at different concentrations.

    Techniques: Neutralization, Infection, Incubation

    Effects of exogenous IFN-β and IL-1α on the expression of type I collagen α1 [Col α1(I)], type III collagen α1 [Col α1(III)], fibronectin (FN), and C/EBPβ mRNA in fibroblasts. The cells were incubated

    Journal:

    Article Title: Host Cell Cytokines Induced by Chlamydia pneumoniae Decrease the Expression of Interstitial Collagens and Fibronectin in Fibroblasts ▿

    doi: 10.1128/IAI.00566-08

    Figure Lengend Snippet: Effects of exogenous IFN-β and IL-1α on the expression of type I collagen α1 [Col α1(I)], type III collagen α1 [Col α1(III)], fibronectin (FN), and C/EBPβ mRNA in fibroblasts. The cells were incubated

    Article Snippet: Fibroblasts grown in six-well plates were stimulated with recombinant IFN-β (PromoCell) or IL-1α (PromoCell) at different concentrations.

    Techniques: Expressing, Incubation

    Experimental evidence that regulation of parkin expression and phosphorylation is related to glial activation and excess IL-1β in Alzheimer brain and in AD Model Systems. a Parkin in neurons in brains of age-matched control patients (AMC) is diffusely distributed within the cytoplasm whereas parkin in neurons in brains of Alzheimer patients (AD) is present in rosette-like aggregates ( b ). c Parkin in Alzheimer brain is colocalized with tau aggregates. d Activated glia overexpressing IL-1α (brown) are immediately adjacent to parkin-immunoreactive neurons (blue), scale bar 20 μM. e Comparative levels of IL-1α, IL-1β, and parkin mRNAs in wild-type littermates or PD-APP mice. f Representative western immunoblot showing proteins from NT2 cells either treated, or not, overnight with IL-1β. g Phospho-parkin levels (P-parkin Ser 65 ) in two representative western immunoblots of proteins from NT2 cell and rat primary neuron cultures treated, or not, with IL-1β. h Quantification of parkin in untreated and treated cell cultures ( n = 6), data is mean ± SEM (** p

    Journal: Journal of Neuroinflammation

    Article Title: Interleukin-1β drives NEDD8 nuclear-to-cytoplasmic translocation, fostering parkin activation via NEDD8 binding to the P-ubiquitin activating site

    doi: 10.1186/s12974-019-1669-z

    Figure Lengend Snippet: Experimental evidence that regulation of parkin expression and phosphorylation is related to glial activation and excess IL-1β in Alzheimer brain and in AD Model Systems. a Parkin in neurons in brains of age-matched control patients (AMC) is diffusely distributed within the cytoplasm whereas parkin in neurons in brains of Alzheimer patients (AD) is present in rosette-like aggregates ( b ). c Parkin in Alzheimer brain is colocalized with tau aggregates. d Activated glia overexpressing IL-1α (brown) are immediately adjacent to parkin-immunoreactive neurons (blue), scale bar 20 μM. e Comparative levels of IL-1α, IL-1β, and parkin mRNAs in wild-type littermates or PD-APP mice. f Representative western immunoblot showing proteins from NT2 cells either treated, or not, overnight with IL-1β. g Phospho-parkin levels (P-parkin Ser 65 ) in two representative western immunoblots of proteins from NT2 cell and rat primary neuron cultures treated, or not, with IL-1β. h Quantification of parkin in untreated and treated cell cultures ( n = 6), data is mean ± SEM (** p

    Article Snippet: For double staining with IL-1α (sc-1279, Santa Cruz Biotechnology) and parkin, endogenous background was blocked with 100% normal horse serum for 30 min at RT.

    Techniques: Expressing, Activation Assay, Mouse Assay, Western Blot

    Effect of TGF-β on dexamethasone (Dex, 10 nM)-induced increases in (A) IκBα, (B) SCNN1A and (C) GILZ mRNA levels after 4 h incubation with IL-1α (1 ng·mL −1 ). Data are presented as the mean and SEM of n = 3 experiments (each determined in triplicate) and the gene expression is normalized to concurrently measured 18 s RNA levels and referenced to levels in control/IL-1α treated cells (1.0). * P

    Journal: British Journal of Pharmacology

    Article Title: Transforming growth factor-? impairs glucocorticoid activity in the A549 lung adenocarcinoma cell line

    doi: 10.1111/j.1476-5381.2012.01885.x

    Figure Lengend Snippet: Effect of TGF-β on dexamethasone (Dex, 10 nM)-induced increases in (A) IκBα, (B) SCNN1A and (C) GILZ mRNA levels after 4 h incubation with IL-1α (1 ng·mL −1 ). Data are presented as the mean and SEM of n = 3 experiments (each determined in triplicate) and the gene expression is normalized to concurrently measured 18 s RNA levels and referenced to levels in control/IL-1α treated cells (1.0). * P

    Article Snippet: Supernatants were collected before treatment with dexamethasone, 24 and 48 h after incubation with IL-1α, and stored at −20°C for the assessment of gene activity by determining the amount of SEAP using a chemiluminescence kit (Roche Applied Science, NSW, Australia) with chemiluminescence measured using Topcount (Perkin Elmer, Glen Waverley, VIC, Australia).

    Techniques: Incubation, Expressing

    Effect of the ALK5 inhibitor (SB431542, 1 µM) on TGF-β-induced glucocorticoid impairment when added 30 min before TGF-β (A) or 30 min prior to IL-1α (B). n = 9 independent experiments. * P

    Journal: British Journal of Pharmacology

    Article Title: Transforming growth factor-? impairs glucocorticoid activity in the A549 lung adenocarcinoma cell line

    doi: 10.1111/j.1476-5381.2012.01885.x

    Figure Lengend Snippet: Effect of the ALK5 inhibitor (SB431542, 1 µM) on TGF-β-induced glucocorticoid impairment when added 30 min before TGF-β (A) or 30 min prior to IL-1α (B). n = 9 independent experiments. * P

    Article Snippet: Supernatants were collected before treatment with dexamethasone, 24 and 48 h after incubation with IL-1α, and stored at −20°C for the assessment of gene activity by determining the amount of SEAP using a chemiluminescence kit (Roche Applied Science, NSW, Australia) with chemiluminescence measured using Topcount (Perkin Elmer, Glen Waverley, VIC, Australia).

    Techniques:

    Characteristics of endogenous intracellular ITEGE neoepitope generated by bovine articular chondrocytes. Bovine articular chondrocytes were released from alginate beads following a 48-hour pretreatment with 10 ng/ml interleukin-1α. A and B , Overlay

    Journal:

    Article Title: The Accumulation of Intracellular ITEGE and DIPEN Neoepitopes in Bovine Articular Chondrocytes Is Mediated by CD44 Internalization of Hyaluronan

    doi: 10.1002/art.21623

    Figure Lengend Snippet: Characteristics of endogenous intracellular ITEGE neoepitope generated by bovine articular chondrocytes. Bovine articular chondrocytes were released from alginate beads following a 48-hour pretreatment with 10 ng/ml interleukin-1α. A and B , Overlay

    Article Snippet: In some experiments the chondrocytes were treated with 10 ng/ml IL-1α (Genzyme, Cambridge, MA) in DMEM containing 10% FBS for 48 hours prior to release from the beads.

    Techniques: Generated

    CM from IL-1α deficient cells fails to support cancer cell invasiveness. The Boyden chamber invasion assay was used to determine the effects of CM from senescent HCA2 cells infected with shGFP or shIL-1α-expressing (#1 and #3) lentiviruses

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Cell surface-bound IL-1α is an upstream regulator of the senescence-associated IL-6/IL-8 cytokine network

    doi: 10.1073/pnas.0905299106

    Figure Lengend Snippet: CM from IL-1α deficient cells fails to support cancer cell invasiveness. The Boyden chamber invasion assay was used to determine the effects of CM from senescent HCA2 cells infected with shGFP or shIL-1α-expressing (#1 and #3) lentiviruses

    Article Snippet: Lentiviruses encoding shRNAs against GFP and IL-1α were purchased from Open Biosystems.

    Techniques: Invasion Assay, Infection, Expressing

    Recombinant IL-1α complements IL-1α deficient senescent cells. ( A ) HCA2 cells infected with shGFP or shIL-1α-expressing (#1 and #3) lentiviruses were treated with bleomycin and allowed to recover for 6 d. Recombinant IL-1α

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Cell surface-bound IL-1α is an upstream regulator of the senescence-associated IL-6/IL-8 cytokine network

    doi: 10.1073/pnas.0905299106

    Figure Lengend Snippet: Recombinant IL-1α complements IL-1α deficient senescent cells. ( A ) HCA2 cells infected with shGFP or shIL-1α-expressing (#1 and #3) lentiviruses were treated with bleomycin and allowed to recover for 6 d. Recombinant IL-1α

    Article Snippet: Lentiviruses encoding shRNAs against GFP and IL-1α were purchased from Open Biosystems.

    Techniques: Recombinant, Infection, Expressing

    Expression and secretion of IL-6 and IL-8 depend on IL-1α signaling. ( A ) HCA2 cells were either untreated or treated with bleomycin and allowed to recover for 6 d. Neutralizing antibodies or rIL-1ra were added for 24 h. CM were collected and analyzed

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Cell surface-bound IL-1α is an upstream regulator of the senescence-associated IL-6/IL-8 cytokine network

    doi: 10.1073/pnas.0905299106

    Figure Lengend Snippet: Expression and secretion of IL-6 and IL-8 depend on IL-1α signaling. ( A ) HCA2 cells were either untreated or treated with bleomycin and allowed to recover for 6 d. Neutralizing antibodies or rIL-1ra were added for 24 h. CM were collected and analyzed

    Article Snippet: Lentiviruses encoding shRNAs against GFP and IL-1α were purchased from Open Biosystems.

    Techniques: Expressing

    IL-1α and IL-1β mRNA and protein levels increase in senescent cells. ( A ) HCA2 fibroblasts were either untreated or treated with bleomycin (50 μg/ml) for 3 h. Total RNA was collected at the indicated intervals thereafter, and transcript

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Cell surface-bound IL-1α is an upstream regulator of the senescence-associated IL-6/IL-8 cytokine network

    doi: 10.1073/pnas.0905299106

    Figure Lengend Snippet: IL-1α and IL-1β mRNA and protein levels increase in senescent cells. ( A ) HCA2 fibroblasts were either untreated or treated with bleomycin (50 μg/ml) for 3 h. Total RNA was collected at the indicated intervals thereafter, and transcript

    Article Snippet: Lentiviruses encoding shRNAs against GFP and IL-1α were purchased from Open Biosystems.

    Techniques:

    Inhibition of IL-1α suppresses the IL-6/IL-8 cytokine response. ( A ) HCA2 cells were infected with shGFP- or shIL-1α-expressing lentiviruses, selected, and expanded. Total mRNA was collected and transcripts for IL-1α and GAPDH were

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Cell surface-bound IL-1α is an upstream regulator of the senescence-associated IL-6/IL-8 cytokine network

    doi: 10.1073/pnas.0905299106

    Figure Lengend Snippet: Inhibition of IL-1α suppresses the IL-6/IL-8 cytokine response. ( A ) HCA2 cells were infected with shGFP- or shIL-1α-expressing lentiviruses, selected, and expanded. Total mRNA was collected and transcripts for IL-1α and GAPDH were

    Article Snippet: Lentiviruses encoding shRNAs against GFP and IL-1α were purchased from Open Biosystems.

    Techniques: Inhibition, Infection, Expressing

    IL-1α regulates IL-6/IL-8 secretion by cells induced to senesce by different stimuli. ( A ) HCA2 cells infected with shGFP or shIL-1α-expressing (#1 and #3) lentiviruses were either untreated or treated with 2 mM NaB for 2 d. CM were collected

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Cell surface-bound IL-1α is an upstream regulator of the senescence-associated IL-6/IL-8 cytokine network

    doi: 10.1073/pnas.0905299106

    Figure Lengend Snippet: IL-1α regulates IL-6/IL-8 secretion by cells induced to senesce by different stimuli. ( A ) HCA2 cells infected with shGFP or shIL-1α-expressing (#1 and #3) lentiviruses were either untreated or treated with 2 mM NaB for 2 d. CM were collected

    Article Snippet: Lentiviruses encoding shRNAs against GFP and IL-1α were purchased from Open Biosystems.

    Techniques: Infection, Expressing

    TRAIL expression and modulation in pIL6-TRAIL + -GFP + -UC-MSCs. a Left: transfection efficiency of UC-MSCs evaluated by flow cytometry for GFP fluorescence. Wildtype UC-MSCs were negative as compared to GFP + -UC-MSCs (red histogram: 83.5%) in a similar amount as for pIL6-TRAIL + -GFP + -UC-MSCs (blue histogram: 87.6%). Right: FACS analysis revealed constitutive TRAIL expression in transduced UC-MSCs (green histogram: 62.5%) that was significantly enhanced by the conditioned medium from U-266 cells (blue histogram: 80.3%) as well as in response to IL-1α/IL-1β (red histogram: 91.2%). b Confocal microscopy of pIL6-TRAIL + -GFP + -UC-MSCs confirmed the upregulation of TRAIL (FITC, green) induced by either U-266 supernatant or IL-1α/IL-1β treatment. Phalloidin staining was used to show actin (TRITC-red); nuclei were counterstained with Hoechst 33342 (blue). Magnification: 100×. c Evaluation of TRAIL expression in transduced cells by RT-qPCR, western blot analysis, and ELISA. (i) After either U-266 or IL-1α/IL-1β treatment, TRAIL mRNA levels in pIL6-TRAIL + -GFP + -UC-MSCs increased up to 2.2-fold and 2.5-fold compared to basal condition. Data are relative amount of mRNA expression normalized to GAPDH and are presented as mean ± SD of three independent experiments. * p

    Journal: Stem Cell Research & Therapy

    Article Title: pIL6-TRAIL-engineered umbilical cord mesenchymal/stromal stem cells are highly cytotoxic for myeloma cells both in vitro and in vivo

    doi: 10.1186/s13287-017-0655-6

    Figure Lengend Snippet: TRAIL expression and modulation in pIL6-TRAIL + -GFP + -UC-MSCs. a Left: transfection efficiency of UC-MSCs evaluated by flow cytometry for GFP fluorescence. Wildtype UC-MSCs were negative as compared to GFP + -UC-MSCs (red histogram: 83.5%) in a similar amount as for pIL6-TRAIL + -GFP + -UC-MSCs (blue histogram: 87.6%). Right: FACS analysis revealed constitutive TRAIL expression in transduced UC-MSCs (green histogram: 62.5%) that was significantly enhanced by the conditioned medium from U-266 cells (blue histogram: 80.3%) as well as in response to IL-1α/IL-1β (red histogram: 91.2%). b Confocal microscopy of pIL6-TRAIL + -GFP + -UC-MSCs confirmed the upregulation of TRAIL (FITC, green) induced by either U-266 supernatant or IL-1α/IL-1β treatment. Phalloidin staining was used to show actin (TRITC-red); nuclei were counterstained with Hoechst 33342 (blue). Magnification: 100×. c Evaluation of TRAIL expression in transduced cells by RT-qPCR, western blot analysis, and ELISA. (i) After either U-266 or IL-1α/IL-1β treatment, TRAIL mRNA levels in pIL6-TRAIL + -GFP + -UC-MSCs increased up to 2.2-fold and 2.5-fold compared to basal condition. Data are relative amount of mRNA expression normalized to GAPDH and are presented as mean ± SD of three independent experiments. * p

    Article Snippet: Finally, soluble TRAIL was also measured in supernatants of pIL6-TRAIL + -GFP + -UC-MSCs after treatment with U-266 conditioned medium, or IL-1α and IL-1β, by dedicated ELISA kit (Abcam).

    Techniques: Expressing, Transfection, Flow Cytometry, Cytometry, Fluorescence, FACS, Confocal Microscopy, Staining, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

    In vitro apoptosis of U-266 cells induced by pIL6-TRAIL + -GFP + -UC-MSCs. a Apoptosis in U-266 cells by transduced UC-MSCs was measured by Annexin V/PI staining using flow cytometry. Representative dot plots revealed that the apoptosis extent was significantly increased (43.8%) after 24 h of coculture with pIL6-TRAIL + -GFP + -UC-MSCs with respect to control UC-MSCs (19.4%) and GFP + -UC-MSCs (20.2%). The effect was also enhanced when IL-1α and IL-1β were added to the cocultures (70.9% of U-266 cell apoptosis). b Active caspase-8 in U-266 cells as signature of TRAIL-induced apoptosis was measured by flow cytometry after 24 h of coculture with UC-MSCs, GFP + -UC-MSCs, and pIL6-TRAIL + -GFP + -UC-MSCs. This representative experiment depicts the activity of caspase-8 in 97.8% of U266 cells cocultured with pIL6-TRAIL + -GFP + -UC-MSCs as compared to 17.9% of control UC-MSCs, and 22% of GFP + -UC-MSCs. Such high levels of active caspase-8 were not further modified by supplementing the cultures with IL-1α/IL-1β (98.9% of positive cells). GFP green fluorescent protein, MSC mesenchymal/stromal stem cell, pIL6 interleukin-6 promoter, TRAIL tumor necrosis factor related apoptosis inducing ligand, UC umbilical cord, WT wildtype

    Journal: Stem Cell Research & Therapy

    Article Title: pIL6-TRAIL-engineered umbilical cord mesenchymal/stromal stem cells are highly cytotoxic for myeloma cells both in vitro and in vivo

    doi: 10.1186/s13287-017-0655-6

    Figure Lengend Snippet: In vitro apoptosis of U-266 cells induced by pIL6-TRAIL + -GFP + -UC-MSCs. a Apoptosis in U-266 cells by transduced UC-MSCs was measured by Annexin V/PI staining using flow cytometry. Representative dot plots revealed that the apoptosis extent was significantly increased (43.8%) after 24 h of coculture with pIL6-TRAIL + -GFP + -UC-MSCs with respect to control UC-MSCs (19.4%) and GFP + -UC-MSCs (20.2%). The effect was also enhanced when IL-1α and IL-1β were added to the cocultures (70.9% of U-266 cell apoptosis). b Active caspase-8 in U-266 cells as signature of TRAIL-induced apoptosis was measured by flow cytometry after 24 h of coculture with UC-MSCs, GFP + -UC-MSCs, and pIL6-TRAIL + -GFP + -UC-MSCs. This representative experiment depicts the activity of caspase-8 in 97.8% of U266 cells cocultured with pIL6-TRAIL + -GFP + -UC-MSCs as compared to 17.9% of control UC-MSCs, and 22% of GFP + -UC-MSCs. Such high levels of active caspase-8 were not further modified by supplementing the cultures with IL-1α/IL-1β (98.9% of positive cells). GFP green fluorescent protein, MSC mesenchymal/stromal stem cell, pIL6 interleukin-6 promoter, TRAIL tumor necrosis factor related apoptosis inducing ligand, UC umbilical cord, WT wildtype

    Article Snippet: Finally, soluble TRAIL was also measured in supernatants of pIL6-TRAIL + -GFP + -UC-MSCs after treatment with U-266 conditioned medium, or IL-1α and IL-1β, by dedicated ELISA kit (Abcam).

    Techniques: In Vitro, Staining, Flow Cytometry, Cytometry, Activity Assay, Modification

    Melatonin inhibits p65 phosphorylation and nuclear translocation in thyroid carcinoma. (A) Western blot analysis of nuclear NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells treated with melatonin for 24 h. (B) ELISA analysis of NF-κB/p65 activity in 8505c, TPC-1 and KTC-1 cells after treatment with melatonin for 24 h. (C) Expression of NF-κB/p65 response genes in 8505c, TPC-1 and KTC-1 cells was detected by qPCR assays. (D) Immunoblotting of IL-1α, Cyclin D1 and MMP9 in 8505c, TPC-1 and KTC-1 cells after treatment with melatonin (2 mM) for 24 h. β-Actin was used as a loading control. (E) Immunoblotting of NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells treated with or without TNFα (10 ng/mL) and melatonin (2 mM) for 24 h. β-Actin was used as a loading control. * P

    Journal: Redox Biology

    Article Title: Melatonin suppresses thyroid cancer growth and overcomes radioresistance via inhibition of p65 phosphorylation and induction of ROS

    doi: 10.1016/j.redox.2018.02.025

    Figure Lengend Snippet: Melatonin inhibits p65 phosphorylation and nuclear translocation in thyroid carcinoma. (A) Western blot analysis of nuclear NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells treated with melatonin for 24 h. (B) ELISA analysis of NF-κB/p65 activity in 8505c, TPC-1 and KTC-1 cells after treatment with melatonin for 24 h. (C) Expression of NF-κB/p65 response genes in 8505c, TPC-1 and KTC-1 cells was detected by qPCR assays. (D) Immunoblotting of IL-1α, Cyclin D1 and MMP9 in 8505c, TPC-1 and KTC-1 cells after treatment with melatonin (2 mM) for 24 h. β-Actin was used as a loading control. (E) Immunoblotting of NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells treated with or without TNFα (10 ng/mL) and melatonin (2 mM) for 24 h. β-Actin was used as a loading control. * P

    Article Snippet: Antibodies used in our study were as follows: β-Actin, PARP, cleaved (c)-caspase 3, NF-κB/p65, p-NF-κB/p65, Bcl-xl, MMP9, Cyclin D1, IL-1α and Lamin A (Cell Signaling Technology, Beverly, MA, USA), Ki-67 (Abcam, Cambridge, Massachusetts, USA).

    Techniques: Translocation Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Activity Assay, Expressing, Real-time Polymerase Chain Reaction

    Irradiation activates NF-κB/p65 phosphorylation. (A) Western blot analysis of nuclear NF-κB/p65 and p-NF-κB/p65 in 8505c, TPC-1 and KTC-1 cells after irradiation. (B) Expression of NF-κB/p65 response genes in 8505c, TPC-1 and KTC-1 cells after irradiation was detected by qPCR assays. (C) Immunoblotting of IL-1α, Cyclin D1 and MMP9 in 8505c, TPC-1 and KTC-1 cells after irradiation. β-Actin was used as a loading control. (D) qPCR analysis of Cyclin D1 and MMP9 in 8505c and TPC-1 cells after treatment with melatonin, irradiation or both. (E) Western blot analysis of nuclear NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells after treated with melatonin, irradiation or both. (F) ELISA analysis of NF-κB/p65 activity in 8505c, TPC-1 and KTC-1 cells after treated with melatonin, irradiation or both. *P

    Journal: Redox Biology

    Article Title: Melatonin suppresses thyroid cancer growth and overcomes radioresistance via inhibition of p65 phosphorylation and induction of ROS

    doi: 10.1016/j.redox.2018.02.025

    Figure Lengend Snippet: Irradiation activates NF-κB/p65 phosphorylation. (A) Western blot analysis of nuclear NF-κB/p65 and p-NF-κB/p65 in 8505c, TPC-1 and KTC-1 cells after irradiation. (B) Expression of NF-κB/p65 response genes in 8505c, TPC-1 and KTC-1 cells after irradiation was detected by qPCR assays. (C) Immunoblotting of IL-1α, Cyclin D1 and MMP9 in 8505c, TPC-1 and KTC-1 cells after irradiation. β-Actin was used as a loading control. (D) qPCR analysis of Cyclin D1 and MMP9 in 8505c and TPC-1 cells after treatment with melatonin, irradiation or both. (E) Western blot analysis of nuclear NF-κB/p65 and p-NF-κB/p65 in 8505c and TPC-1 cells after treated with melatonin, irradiation or both. (F) ELISA analysis of NF-κB/p65 activity in 8505c, TPC-1 and KTC-1 cells after treated with melatonin, irradiation or both. *P

    Article Snippet: Antibodies used in our study were as follows: β-Actin, PARP, cleaved (c)-caspase 3, NF-κB/p65, p-NF-κB/p65, Bcl-xl, MMP9, Cyclin D1, IL-1α and Lamin A (Cell Signaling Technology, Beverly, MA, USA), Ki-67 (Abcam, Cambridge, Massachusetts, USA).

    Techniques: Irradiation, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Activity Assay

    Spa spring water impairs pro-inflammatory cytokine production on HaCaT cells under TLR stimulation. The HaCaT cells were treated with TLR1 through TLR6 agonist for 1, 4, 10, and 24 hours, respectively. Spa spring water diluted 1 : 30 fold was added prior to adding TLR agonists or simultaneously. After the indicated time, the supernatants were collected to measure secreted level of IL-6 (A), IL-8 (B), TNF-α (C), IL-1α (D), and GM-CSF (E) via ELISA. The data are expressed as the means±SEM ( * p

    Journal: Annals of Dermatology

    Article Title: Effect of Spa Spring Water on Cytokine Expression in Human Keratinocyte HaCaT Cells and on Differentiation of CD4+ T Cells

    doi: 10.5021/ad.2012.24.3.324

    Figure Lengend Snippet: Spa spring water impairs pro-inflammatory cytokine production on HaCaT cells under TLR stimulation. The HaCaT cells were treated with TLR1 through TLR6 agonist for 1, 4, 10, and 24 hours, respectively. Spa spring water diluted 1 : 30 fold was added prior to adding TLR agonists or simultaneously. After the indicated time, the supernatants were collected to measure secreted level of IL-6 (A), IL-8 (B), TNF-α (C), IL-1α (D), and GM-CSF (E) via ELISA. The data are expressed as the means±SEM ( * p

    Article Snippet: ELISA Levels of IL-6, IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α (BD OptEIA™, BD Biosciences Pharmingen, San Diego, CA, USA), IL-1α (Biolegend, San Diego, CA, USA) in HaCaT supernatant treated with TLR agonist in the presence or absence of hot spring water were quantified according to the manufacturer's protocol.

    Techniques: Enzyme-linked Immunosorbent Assay

    IL-1α signals through IL-1R to induce the SASP. (A) Western blot analysis of IL-1α, IL-1β, and tubulin (Tub) levels in IMR90T cells expressing either a doxycycline-inducible scramble shRNA (shScr) or shRNA against IL-1α (shIL1α). Cells were treated with either ethanol (Et) as a control or tamoxifen (4OHT) to induce Ras activity and doxycycline to induce knockdown. (B) qRT-PCR analyses of the indicated genes in the indicated cells. Values are shown as fold change compared to levels in shScr treated with Et. (C) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (D) Western blot analysis detecting IL-1α and Tub levels in wild-type (WT) and IL-1α-deleted (KO) IMR90T cells. WT1, parental cell line; WT2, a clone that failed to delete IL-1α; KO pool, total pool of clones, where KO1 and KO2 each represent an independent clone. (E) qRT-PCR detecting IL-8 mRNA levels in the indicated cells. (F) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (G) qRT-PCR analyses of the indicated SASP factors found via transcriptome analysis to be affected by IL-1R knockdown. Data are represented as fold change compared to levels for WT1 treated with Et. (H) Western blot analysis of IL-1α in IMR90T cells expressing an empty vector control (EV), full-length IL-1α (IL-1α FL), the N-terminal propiece (ppIL-1α), or the C-terminal mature fragment (mIL-1α). The antibody used only recognizes the C-terminal portion of IL-1α. (I) qRT-PCR analyses of the indicated genes in the indicated cell lines. Primers for IL-1α anneal to the N terminus. Values are shown as fold change compared to levels in cells expressing EV. Each experiment was performed at least 3 times. ns, not significant; *, P

    Journal: Molecular and Cellular Biology

    Article Title: Uncoupling the Senescence-Associated Secretory Phenotype from Cell Cycle Exit via Interleukin-1 Inactivation Unveils Its Protumorigenic Role

    doi: 10.1128/MCB.00586-18

    Figure Lengend Snippet: IL-1α signals through IL-1R to induce the SASP. (A) Western blot analysis of IL-1α, IL-1β, and tubulin (Tub) levels in IMR90T cells expressing either a doxycycline-inducible scramble shRNA (shScr) or shRNA against IL-1α (shIL1α). Cells were treated with either ethanol (Et) as a control or tamoxifen (4OHT) to induce Ras activity and doxycycline to induce knockdown. (B) qRT-PCR analyses of the indicated genes in the indicated cells. Values are shown as fold change compared to levels in shScr treated with Et. (C) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (D) Western blot analysis detecting IL-1α and Tub levels in wild-type (WT) and IL-1α-deleted (KO) IMR90T cells. WT1, parental cell line; WT2, a clone that failed to delete IL-1α; KO pool, total pool of clones, where KO1 and KO2 each represent an independent clone. (E) qRT-PCR detecting IL-8 mRNA levels in the indicated cells. (F) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (G) qRT-PCR analyses of the indicated SASP factors found via transcriptome analysis to be affected by IL-1R knockdown. Data are represented as fold change compared to levels for WT1 treated with Et. (H) Western blot analysis of IL-1α in IMR90T cells expressing an empty vector control (EV), full-length IL-1α (IL-1α FL), the N-terminal propiece (ppIL-1α), or the C-terminal mature fragment (mIL-1α). The antibody used only recognizes the C-terminal portion of IL-1α. (I) qRT-PCR analyses of the indicated genes in the indicated cell lines. Primers for IL-1α anneal to the N terminus. Values are shown as fold change compared to levels in cells expressing EV. Each experiment was performed at least 3 times. ns, not significant; *, P

    Article Snippet: To induce IL-1α (no. V2THS_111503; Dharmacon) or IL-1R (no. V2THS_131081 and V2THS_131083; Dharmacon) knockdown, cells were cultured in the presence of 0.5 µg/ml doxycycline (Sigma) for 24 h before the start of senescence induction.

    Techniques: Western Blot, Expressing, shRNA, Activity Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Clone Assay, Plasmid Preparation

    Model of senescence-induced SASP activation under physiological conditions (A) or conditions where either IL-1α or IL-1β is depleted (B).

    Journal: Molecular and Cellular Biology

    Article Title: Uncoupling the Senescence-Associated Secretory Phenotype from Cell Cycle Exit via Interleukin-1 Inactivation Unveils Its Protumorigenic Role

    doi: 10.1128/MCB.00586-18

    Figure Lengend Snippet: Model of senescence-induced SASP activation under physiological conditions (A) or conditions where either IL-1α or IL-1β is depleted (B).

    Article Snippet: To induce IL-1α (no. V2THS_111503; Dharmacon) or IL-1R (no. V2THS_131081 and V2THS_131083; Dharmacon) knockdown, cells were cultured in the presence of 0.5 µg/ml doxycycline (Sigma) for 24 h before the start of senescence induction.

    Techniques: Activation Assay

    IL-1α can induce the SASP in the absence of IL-1β and vice versa. (A) Western blot analysis of IL-1α, IL-1β, and tubulin (Tub) levels in IMR90T cells expressing either an empty vector (EV), full-length IL-1α, or myc-tagged mature IL-1α (mIL-1α). IMR90T cells were also expressing either a scramble shRNA (shScr), a doxycycline-inducible shRNA against IL-1R (shIL-1R), or a constitutively active shRNA against IL-1β (shIL-1β). (B) mRNA expression levels of the indicated genes in the indicated cell lines as detected by qRT-PCR. Values are expressed as fold change compared to levels for shScr cells expressing EV. (C) Western blot analysis of IL-1β and Tub levels in IMR90T cells expressing either EV, full-length IL-1β, or mature IL-1β (mIL-1β). Cells were also expressing either a doxycycline-inducible scramble shRNA (shScr) or a doxycycline-inducible shRNA against IL-1R (shIL-1R) or IL-1α (shIL-1α). (D) mRNA expression levels of the indicated genes in the indicated cell lines as detected by qRT-PCR. Values are expressed as fold change compared to levels in shScr cells expressing EV. Each experiment was repeated at least 3 times. *, P

    Journal: Molecular and Cellular Biology

    Article Title: Uncoupling the Senescence-Associated Secretory Phenotype from Cell Cycle Exit via Interleukin-1 Inactivation Unveils Its Protumorigenic Role

    doi: 10.1128/MCB.00586-18

    Figure Lengend Snippet: IL-1α can induce the SASP in the absence of IL-1β and vice versa. (A) Western blot analysis of IL-1α, IL-1β, and tubulin (Tub) levels in IMR90T cells expressing either an empty vector (EV), full-length IL-1α, or myc-tagged mature IL-1α (mIL-1α). IMR90T cells were also expressing either a scramble shRNA (shScr), a doxycycline-inducible shRNA against IL-1R (shIL-1R), or a constitutively active shRNA against IL-1β (shIL-1β). (B) mRNA expression levels of the indicated genes in the indicated cell lines as detected by qRT-PCR. Values are expressed as fold change compared to levels for shScr cells expressing EV. (C) Western blot analysis of IL-1β and Tub levels in IMR90T cells expressing either EV, full-length IL-1β, or mature IL-1β (mIL-1β). Cells were also expressing either a doxycycline-inducible scramble shRNA (shScr) or a doxycycline-inducible shRNA against IL-1R (shIL-1R) or IL-1α (shIL-1α). (D) mRNA expression levels of the indicated genes in the indicated cell lines as detected by qRT-PCR. Values are expressed as fold change compared to levels in shScr cells expressing EV. Each experiment was repeated at least 3 times. *, P

    Article Snippet: To induce IL-1α (no. V2THS_111503; Dharmacon) or IL-1R (no. V2THS_131081 and V2THS_131083; Dharmacon) knockdown, cells were cultured in the presence of 0.5 µg/ml doxycycline (Sigma) for 24 h before the start of senescence induction.

    Techniques: Western Blot, Expressing, Plasmid Preparation, shRNA, Quantitative RT-PCR

    IL-1α can be used to uncouple the SASP from cell cycle exit. (A) Quantitation of BrdU incorporation and SA-βgal positivity in IMR90T cells after introduction of an empty vector (EV) or a vector containing full-length IL-1α. (B to D) Representative images and quantitation of BrdU incorporation (B), SA-βgal positivity (C), and SAHF positivity (D) in the indicated cell lines. Scale bars for panels B and C, 200 µm; scale bar for panel D, 12 µm. Each experiment was performed at least 3 times. ns, not significant; *, P

    Journal: Molecular and Cellular Biology

    Article Title: Uncoupling the Senescence-Associated Secretory Phenotype from Cell Cycle Exit via Interleukin-1 Inactivation Unveils Its Protumorigenic Role

    doi: 10.1128/MCB.00586-18

    Figure Lengend Snippet: IL-1α can be used to uncouple the SASP from cell cycle exit. (A) Quantitation of BrdU incorporation and SA-βgal positivity in IMR90T cells after introduction of an empty vector (EV) or a vector containing full-length IL-1α. (B to D) Representative images and quantitation of BrdU incorporation (B), SA-βgal positivity (C), and SAHF positivity (D) in the indicated cell lines. Scale bars for panels B and C, 200 µm; scale bar for panel D, 12 µm. Each experiment was performed at least 3 times. ns, not significant; *, P

    Article Snippet: To induce IL-1α (no. V2THS_111503; Dharmacon) or IL-1R (no. V2THS_131081 and V2THS_131083; Dharmacon) knockdown, cells were cultured in the presence of 0.5 µg/ml doxycycline (Sigma) for 24 h before the start of senescence induction.

    Techniques: Quantitation Assay, BrdU Incorporation Assay, Plasmid Preparation

    IL-1α deletion delays pancreatic cancer progression. (A) Schematic of crosses in the mouse model used in this study. Mice containing a Lox-Stop-Lox K-Ras G12D allele were crossed with mice containing a p48-Cre allele to create KC WT mice. KC mice were crossed to mice that have both IL-1α alleles disrupted to create KC IL-1α −/− mice. (B) Representative H E images of pancreata and quantitation of neoplastic area from 12-week-old KC WT and KC IL-1α −/− mice. Scale bar, 200 µm. (C) Representative images and quantitation of Masson trichrome staining. Scale bar, 200 µm. (D) Representative images and quantitation of CD3 staining in total pancreas tissue in the indicated samples. Scale bar, 50 µm. (E) Quantitation of CD3 staining within lesions in the indicated samples. (F and G) Representative images and quantitation of F4/80 staining in total pancreas tissue (F) and within lesions in the indicated samples (G). Scale bars, 200 µm in panel F and 50 µm in panel G. (H) SA-βgal positivity and quantitation in the indicated samples. Scale bar, 200 µm. (I) Ki-67 staining and quantitation in the indicated samples. Scale bar, 50 µm. (J) mRNA expression levels of the indicated genes in the indicated samples. n was at least 10 in panels B to G and J, and n was 5 in panels H and I. ns, not significant; *, P

    Journal: Molecular and Cellular Biology

    Article Title: Uncoupling the Senescence-Associated Secretory Phenotype from Cell Cycle Exit via Interleukin-1 Inactivation Unveils Its Protumorigenic Role

    doi: 10.1128/MCB.00586-18

    Figure Lengend Snippet: IL-1α deletion delays pancreatic cancer progression. (A) Schematic of crosses in the mouse model used in this study. Mice containing a Lox-Stop-Lox K-Ras G12D allele were crossed with mice containing a p48-Cre allele to create KC WT mice. KC mice were crossed to mice that have both IL-1α alleles disrupted to create KC IL-1α −/− mice. (B) Representative H E images of pancreata and quantitation of neoplastic area from 12-week-old KC WT and KC IL-1α −/− mice. Scale bar, 200 µm. (C) Representative images and quantitation of Masson trichrome staining. Scale bar, 200 µm. (D) Representative images and quantitation of CD3 staining in total pancreas tissue in the indicated samples. Scale bar, 50 µm. (E) Quantitation of CD3 staining within lesions in the indicated samples. (F and G) Representative images and quantitation of F4/80 staining in total pancreas tissue (F) and within lesions in the indicated samples (G). Scale bars, 200 µm in panel F and 50 µm in panel G. (H) SA-βgal positivity and quantitation in the indicated samples. Scale bar, 200 µm. (I) Ki-67 staining and quantitation in the indicated samples. Scale bar, 50 µm. (J) mRNA expression levels of the indicated genes in the indicated samples. n was at least 10 in panels B to G and J, and n was 5 in panels H and I. ns, not significant; *, P

    Article Snippet: To induce IL-1α (no. V2THS_111503; Dharmacon) or IL-1R (no. V2THS_131081 and V2THS_131083; Dharmacon) knockdown, cells were cultured in the presence of 0.5 µg/ml doxycycline (Sigma) for 24 h before the start of senescence induction.

    Techniques: Mouse Assay, Quantitation Assay, Staining, Expressing

    IL-1β is also required for SASP expression but not cell cycle exit. (A) Western blot analysis of IL-1β, IL-1α, and tubulin (Tub) levels in IMR90T cells expressing either an empty vector (EV) or shRNA against IL-1β (shIL1β). Cells were treated with either ethanol (Et) as a control or tamoxifen (4OHT) to induce Ras activity. (B) mRNA expression levels of the indicated genes in the indicated cells as detected by qRT-PCR. Values are shown as fold change compared to the level for EV treated with Et. (C) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (D to F) Representative images and quantitation of BrdU incorporation (D), SA-βgal positivity (E), and SAHF positivity (F) in the indicated cell lines. Scale bars for panels D and E, 200 µm; scale bar for panel F, 12 µm. Each experiment was performed at least 3 times. *, P

    Journal: Molecular and Cellular Biology

    Article Title: Uncoupling the Senescence-Associated Secretory Phenotype from Cell Cycle Exit via Interleukin-1 Inactivation Unveils Its Protumorigenic Role

    doi: 10.1128/MCB.00586-18

    Figure Lengend Snippet: IL-1β is also required for SASP expression but not cell cycle exit. (A) Western blot analysis of IL-1β, IL-1α, and tubulin (Tub) levels in IMR90T cells expressing either an empty vector (EV) or shRNA against IL-1β (shIL1β). Cells were treated with either ethanol (Et) as a control or tamoxifen (4OHT) to induce Ras activity. (B) mRNA expression levels of the indicated genes in the indicated cells as detected by qRT-PCR. Values are shown as fold change compared to the level for EV treated with Et. (C) Secreted levels of IL-6 in the indicated cell lines as detected by ELISA. (D to F) Representative images and quantitation of BrdU incorporation (D), SA-βgal positivity (E), and SAHF positivity (F) in the indicated cell lines. Scale bars for panels D and E, 200 µm; scale bar for panel F, 12 µm. Each experiment was performed at least 3 times. *, P

    Article Snippet: To induce IL-1α (no. V2THS_111503; Dharmacon) or IL-1R (no. V2THS_131081 and V2THS_131083; Dharmacon) knockdown, cells were cultured in the presence of 0.5 µg/ml doxycycline (Sigma) for 24 h before the start of senescence induction.

    Techniques: Expressing, Western Blot, Plasmid Preparation, shRNA, Activity Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Quantitation Assay, BrdU Incorporation Assay

    CD11a hi FcγRIII hi B cells produce a high level of IFN-γ in response to CD40 ligation. C57BL/6 mice were infected with 2 × 10 6 LM. (A) The intracellular expression of IL-1α, IL-2, IL-6, IL-10 and IFN-γ in CD11a hi FcγRIII hi and conventional B cells was assayed on day 3 post-infection. (B - G) Splenic CD11a hi FcγRIII hi and conventional B cells were sorted on day 3 post-infection. IL-1β, IL-2, IL-6, IL-12p70, IFN-γ, or TNF-α secretion by CD11a hi FcγRIII hi and conventional B cells was detected by ELISA. Data shown represent the mean ± SD of triplicate experiments. * P

    Journal: Cell Research

    Article Title: Identification of IFN-?-producing innate B cells

    doi: 10.1038/cr.2013.155

    Figure Lengend Snippet: CD11a hi FcγRIII hi B cells produce a high level of IFN-γ in response to CD40 ligation. C57BL/6 mice were infected with 2 × 10 6 LM. (A) The intracellular expression of IL-1α, IL-2, IL-6, IL-10 and IFN-γ in CD11a hi FcγRIII hi and conventional B cells was assayed on day 3 post-infection. (B - G) Splenic CD11a hi FcγRIII hi and conventional B cells were sorted on day 3 post-infection. IL-1β, IL-2, IL-6, IL-12p70, IFN-γ, or TNF-α secretion by CD11a hi FcγRIII hi and conventional B cells was detected by ELISA. Data shown represent the mean ± SD of triplicate experiments. * P

    Article Snippet: Fluorescence-conjugated mAbs against CD4 (RM4-5), CD5 (53-7.3), CD8 (53-6.7), CD11a (2D7), CD11b (M1/70), CD11c (HL3), CD16/32 (2.4G2), CD19 (1D3), CD21/CD35 (7G6), CD23 (B3B4), CD40 (3/23), CD40L (TRAP1), CD43 (S7), CD45R/B220 (RA3-6B2), CD80 (16-10A1), CD86 (GL1), CD93 (C1qRp) (R139), F4/80 (6F12), NK1.1 (PK136), I-A/I-E (2G9), IgD (11-26c.2a), IgM (G155-228), IL-1α (364-3B3-14), IL-2 (MQ1-17H12), IL-6 (MP5-20F3), IL-10 (JES3-19F1), IFN-γ (B27), IgD (11-26c.2a) and IgM (G155-228) and agonistic anti-CD40 (HM40-3) and neutralizing CD11a mAbs (M17/4) were purchased from BD Biosciences Pharmingen (San Diego, CA, USA).

    Techniques: Ligation, Mouse Assay, Infection, Expressing, Enzyme-linked Immunosorbent Assay

    Effect of IL-1α, IL-1α antibody and IL-1R1 antagonist on the expression of IL-1R1 and S100A9 mRNA in HaCaT cells

    Journal: Biochimica et biophysica acta

    Article Title: Mechanism of interleukin-1α transcriptional regulation of S100A9 in a human epidermal keratinocyte cell line

    doi: 10.1016/j.bbagrm.2013.03.010

    Figure Lengend Snippet: Effect of IL-1α, IL-1α antibody and IL-1R1 antagonist on the expression of IL-1R1 and S100A9 mRNA in HaCaT cells

    Article Snippet: At 90% confluence, HaCaT cells were further cultured with or without IL-1α (20 ng/ml; Wako Pure Chemical Industries, Osaka, Japan) for 24 h. In the experiments using inhibitors, cells were cultured in DMEM-2% FBS for 12 h and then pre-cultured for 1 h with anti-human IL-1α antibody (1/100 dilution; IgG fraction of anti-human IL-1α, Rockland, Gilbertsville, PA, USA), IL-1α receptor antagonist (100 ng/ml; recombinant human IL-1ra/IL-1F3; IL-1ra, R & D SYSTEMS, Minneapolis, USA) or MAPK inhibitors dissolved in DMSO (included in buffer controls as appropriate) including SB203580 (30 μM; Wako), U0126 (10 μM; Wako) or SP600125 (10 μM; CALBIOCHEM, Darmstadt, Germany), and cultured with IL-1α (20 ng/ml) for 30 min (Western blotting for MAP kinase phosphorylation) or 24 h (gene expression studies using Northern blotting and quantitative PCR).

    Techniques: Expressing

    Effect of IL-1α on S100A9 transcription in HaCaT cells

    Journal: Biochimica et biophysica acta

    Article Title: Mechanism of interleukin-1α transcriptional regulation of S100A9 in a human epidermal keratinocyte cell line

    doi: 10.1016/j.bbagrm.2013.03.010

    Figure Lengend Snippet: Effect of IL-1α on S100A9 transcription in HaCaT cells

    Article Snippet: At 90% confluence, HaCaT cells were further cultured with or without IL-1α (20 ng/ml; Wako Pure Chemical Industries, Osaka, Japan) for 24 h. In the experiments using inhibitors, cells were cultured in DMEM-2% FBS for 12 h and then pre-cultured for 1 h with anti-human IL-1α antibody (1/100 dilution; IgG fraction of anti-human IL-1α, Rockland, Gilbertsville, PA, USA), IL-1α receptor antagonist (100 ng/ml; recombinant human IL-1ra/IL-1F3; IL-1ra, R & D SYSTEMS, Minneapolis, USA) or MAPK inhibitors dissolved in DMSO (included in buffer controls as appropriate) including SB203580 (30 μM; Wako), U0126 (10 μM; Wako) or SP600125 (10 μM; CALBIOCHEM, Darmstadt, Germany), and cultured with IL-1α (20 ng/ml) for 30 min (Western blotting for MAP kinase phosphorylation) or 24 h (gene expression studies using Northern blotting and quantitative PCR).

    Techniques:

    Secreted MMP activity of STEs and cartilage explants cultured alone or together. After 7 days, the MMP-activity (rfu/sec) was determined (in culture medium) of normal ( A, B ) and OA ( C, D ) STEs and healthy cartilage explants, cultured alone or together, without ( A, C ) or with ( B, D ) IL-1α stimulation. Each line represents an individual donor and demonstrates the activity of STEs alone (left), STEs together with cartilage explants (middle) or cartilage explants cultured alone (right). The MMP activity in the medium of STEs cultured alone was higher, compared to the MMP activity in the co-culture conditions. Significance levels are indicated.

    Journal: PLoS ONE

    Article Title: Profiling the Secretion of Soluble Mediators by End Stage Osteoarthritis Synovial Tissue Explants Reveals a Reduced Responsiveness to an Inflammatory Trigger

    doi: 10.1371/journal.pone.0062634

    Figure Lengend Snippet: Secreted MMP activity of STEs and cartilage explants cultured alone or together. After 7 days, the MMP-activity (rfu/sec) was determined (in culture medium) of normal ( A, B ) and OA ( C, D ) STEs and healthy cartilage explants, cultured alone or together, without ( A, C ) or with ( B, D ) IL-1α stimulation. Each line represents an individual donor and demonstrates the activity of STEs alone (left), STEs together with cartilage explants (middle) or cartilage explants cultured alone (right). The MMP activity in the medium of STEs cultured alone was higher, compared to the MMP activity in the co-culture conditions. Significance levels are indicated.

    Article Snippet: STEs Culture As a reflection of the whole synovium, for each condition 6 STEs from different anatomical locations in the joint per donor were cultured in a 24 wells plate in 700 µl Dulbecco’s Modified Eagle Medium (DMEM) (Invitrogen) supplemented with Insulin-transferrin-sodium selenite (ITS) (Roche Diagnostics, Basel, Switzerland), 1% v/v Penstrep, 1 mg/ml lactalbumin and 5 µg/ml vitamin C (Sigma-Aldrich) (serum free medium) and were incubated with or without 10 ng/ml IL-1α (Peprotech, Rocky Hill, USA).

    Techniques: Activity Assay, Cell Culture, Size-exclusion Chromatography, Co-Culture Assay

    Proteoglycan degradation of healthy cartilage explants cultured alone or together with synovial tissue explants (STEs). Proteoglycan degradation was expressed as the percentage of glycosaminoglycans (GAG) released into the medium during 7 days of culture of cartilage alone or co-cultured together with normal ( A, B ) or OA ( C, D ) STEs without ( A, C ) or with ( B, D ) IL-1α stimulation. Each line represents an individual donor and connects the % GAG release of cartilage alone (left) with the matching co-culture condition of cartilage together with STEs (right). There were no significant differences between cartilage cultured alone or co-cultured with normal or OA STEs.

    Journal: PLoS ONE

    Article Title: Profiling the Secretion of Soluble Mediators by End Stage Osteoarthritis Synovial Tissue Explants Reveals a Reduced Responsiveness to an Inflammatory Trigger

    doi: 10.1371/journal.pone.0062634

    Figure Lengend Snippet: Proteoglycan degradation of healthy cartilage explants cultured alone or together with synovial tissue explants (STEs). Proteoglycan degradation was expressed as the percentage of glycosaminoglycans (GAG) released into the medium during 7 days of culture of cartilage alone or co-cultured together with normal ( A, B ) or OA ( C, D ) STEs without ( A, C ) or with ( B, D ) IL-1α stimulation. Each line represents an individual donor and connects the % GAG release of cartilage alone (left) with the matching co-culture condition of cartilage together with STEs (right). There were no significant differences between cartilage cultured alone or co-cultured with normal or OA STEs.

    Article Snippet: STEs Culture As a reflection of the whole synovium, for each condition 6 STEs from different anatomical locations in the joint per donor were cultured in a 24 wells plate in 700 µl Dulbecco’s Modified Eagle Medium (DMEM) (Invitrogen) supplemented with Insulin-transferrin-sodium selenite (ITS) (Roche Diagnostics, Basel, Switzerland), 1% v/v Penstrep, 1 mg/ml lactalbumin and 5 µg/ml vitamin C (Sigma-Aldrich) (serum free medium) and were incubated with or without 10 ng/ml IL-1α (Peprotech, Rocky Hill, USA).

    Techniques: Cell Culture, Co-Culture Assay

    Soluble mediator secretion by normal and OA synovial tissue explants (STEs) with or without IL-1α. Graphs demonstrate all soluble mediators which were significantly (p

    Journal: PLoS ONE

    Article Title: Profiling the Secretion of Soluble Mediators by End Stage Osteoarthritis Synovial Tissue Explants Reveals a Reduced Responsiveness to an Inflammatory Trigger

    doi: 10.1371/journal.pone.0062634

    Figure Lengend Snippet: Soluble mediator secretion by normal and OA synovial tissue explants (STEs) with or without IL-1α. Graphs demonstrate all soluble mediators which were significantly (p

    Article Snippet: STEs Culture As a reflection of the whole synovium, for each condition 6 STEs from different anatomical locations in the joint per donor were cultured in a 24 wells plate in 700 µl Dulbecco’s Modified Eagle Medium (DMEM) (Invitrogen) supplemented with Insulin-transferrin-sodium selenite (ITS) (Roche Diagnostics, Basel, Switzerland), 1% v/v Penstrep, 1 mg/ml lactalbumin and 5 µg/ml vitamin C (Sigma-Aldrich) (serum free medium) and were incubated with or without 10 ng/ml IL-1α (Peprotech, Rocky Hill, USA).

    Techniques:

    IL-1α localization and knockdown in THP-1 macrophages. A ) THP-1 macrophages were transfected with control (ctr) or IL-1α siRNA for 48 h, followed by 24 h stimulation with MSU (100 µg/ml). Cell lysates were used to examine the expression of pro-IL-1α, pro-IL-1β, cleaved capase-1, and NLRP3 in IL-1α knockdown cells. B ) THP-1 macrophages were stimulated with MSU (100 μg/ml) for 30 min. Nuclear and cyotsolic fractions were prepared to evaluate the expression of pro–IL-1α and –IL-1β. C ) Conditioned medium was collected and used for the quantification of IL-1β, TNF-α, IL-8, and MCP-1 production with without MSU stimulation. NS, nonstimulated. * P

    Journal: The FASEB Journal

    Article Title: Critical role of IL-1α in IL-1β–induced inflammatory responses: cooperation with NF-κBp65 in transcriptional regulation

    doi: 10.1096/fj.201801513R

    Figure Lengend Snippet: IL-1α localization and knockdown in THP-1 macrophages. A ) THP-1 macrophages were transfected with control (ctr) or IL-1α siRNA for 48 h, followed by 24 h stimulation with MSU (100 µg/ml). Cell lysates were used to examine the expression of pro-IL-1α, pro-IL-1β, cleaved capase-1, and NLRP3 in IL-1α knockdown cells. B ) THP-1 macrophages were stimulated with MSU (100 μg/ml) for 30 min. Nuclear and cyotsolic fractions were prepared to evaluate the expression of pro–IL-1α and –IL-1β. C ) Conditioned medium was collected and used for the quantification of IL-1β, TNF-α, IL-8, and MCP-1 production with without MSU stimulation. NS, nonstimulated. * P

    Article Snippet: Small interfering RNA (siRNA) sequences for IL-1α were purchased from MilliporeSigma (eMission predesigned siRNA sequences SASI_Hs01_00167953 and SASI_Hs02_00302829).

    Techniques: Transfection, Expressing

    NF-κB governs IL-1α and IL-1β expression. A ) RASFs were pretreated with SB202190 (SB, 10 µM), SP600125 (SP, 10 µM), PD98059 (PD, 10 µM), PDTC (100 µM), or diacerein (DIC, 10 µM) for 2 h prior to IL-1β stimulation for 24 h. Cell lysates were analyzed for IL-1α and IL-1β expression. B ) RASFs (8-well chamber slides) were pretreated with IL-1R antagonist (100 ng/ml) for 2 h followed by IL-1β stimulation for 8 h. C ) Similarly, RASFs were treated with PDTC (100 µM) for 2 h followed by IL-1β stimulation for 8 h. Cells were fixed and analyzed for IL-1α expression using immunofluorescence method. D , E ) Conditioned media from inhibitors study in A were used for determining IL-6 and IL-8 using ELISA. PDTC inhibits both IL-1α and IL-1β, as well as IL-6 and IL-8 production. Cells were lysed and analyzed for IL-1α and IL-1β. * P

    Journal: The FASEB Journal

    Article Title: Critical role of IL-1α in IL-1β–induced inflammatory responses: cooperation with NF-κBp65 in transcriptional regulation

    doi: 10.1096/fj.201801513R

    Figure Lengend Snippet: NF-κB governs IL-1α and IL-1β expression. A ) RASFs were pretreated with SB202190 (SB, 10 µM), SP600125 (SP, 10 µM), PD98059 (PD, 10 µM), PDTC (100 µM), or diacerein (DIC, 10 µM) for 2 h prior to IL-1β stimulation for 24 h. Cell lysates were analyzed for IL-1α and IL-1β expression. B ) RASFs (8-well chamber slides) were pretreated with IL-1R antagonist (100 ng/ml) for 2 h followed by IL-1β stimulation for 8 h. C ) Similarly, RASFs were treated with PDTC (100 µM) for 2 h followed by IL-1β stimulation for 8 h. Cells were fixed and analyzed for IL-1α expression using immunofluorescence method. D , E ) Conditioned media from inhibitors study in A were used for determining IL-6 and IL-8 using ELISA. PDTC inhibits both IL-1α and IL-1β, as well as IL-6 and IL-8 production. Cells were lysed and analyzed for IL-1α and IL-1β. * P

    Article Snippet: Small interfering RNA (siRNA) sequences for IL-1α were purchased from MilliporeSigma (eMission predesigned siRNA sequences SASI_Hs01_00167953 and SASI_Hs02_00302829).

    Techniques: Expressing, Immunofluorescence, Enzyme-linked Immunosorbent Assay

    A ) The interacting surface of the conformation of IL-1R (green) and IL-1β (gray) complex at 100 ns molecular dynamics simulation. The electrostatic potential map of interacting surface of receptor shown in red, blue, and white represent electronegative, electropositive, and hydrophobic surface area of the binding site, respectively. The H-bonds and π-π interaction are shown as black and cyan dotted line, respectively. B ) The interacting surface of the conformation of IL-1R (green) and IL-1α (gray) complex at 100 ns molecular dynamics simulation. The electrostatic potential map of interacting surface of receptor shown in red, blue, and white represent electronegative, electropositive, and hydrophobic surface area of the binding site, respectively. The H-bonds and cation-π interaction are shown as black and green dotted line, respectively.

    Journal: The FASEB Journal

    Article Title: Critical role of IL-1α in IL-1β–induced inflammatory responses: cooperation with NF-κBp65 in transcriptional regulation

    doi: 10.1096/fj.201801513R

    Figure Lengend Snippet: A ) The interacting surface of the conformation of IL-1R (green) and IL-1β (gray) complex at 100 ns molecular dynamics simulation. The electrostatic potential map of interacting surface of receptor shown in red, blue, and white represent electronegative, electropositive, and hydrophobic surface area of the binding site, respectively. The H-bonds and π-π interaction are shown as black and cyan dotted line, respectively. B ) The interacting surface of the conformation of IL-1R (green) and IL-1α (gray) complex at 100 ns molecular dynamics simulation. The electrostatic potential map of interacting surface of receptor shown in red, blue, and white represent electronegative, electropositive, and hydrophobic surface area of the binding site, respectively. The H-bonds and cation-π interaction are shown as black and green dotted line, respectively.

    Article Snippet: Small interfering RNA (siRNA) sequences for IL-1α were purchased from MilliporeSigma (eMission predesigned siRNA sequences SASI_Hs01_00167953 and SASI_Hs02_00302829).

    Techniques: Binding Assay

    Effects of IL-1α knockdown. A ) IL-1α expression was knocked down using IL-1α siRNA for 48 h prior to IL-1β stimulation for 24 h. We observed that knocking down IL-1α also reduced IL-1β expression. B ) We reconfirmed the knockdown of IL-1α using an IL-1α ELISA and stimulating RASFs with ATP (5 nM) and LPS (10 μg/ml) to trigger IL-1α release into conditioned medium. C ) Upon confirming IL-1α has been knocked down, we observed a statistically significant decrease of IL-6 production upon IL-1β stimulation. D ) We also observed a decrease in IL-8 production; however, this was not statistically significant. E ) In addition to released proteins, we analyzed expression of cyclooxygenases 1 and 2 (COX-1 and -2) and saw a statistically significant reduction in IL-1β–induced COX-2 and intercellular adhesion molecule 1 expression in IL-1α knockdown samples. * P

    Journal: The FASEB Journal

    Article Title: Critical role of IL-1α in IL-1β–induced inflammatory responses: cooperation with NF-κBp65 in transcriptional regulation

    doi: 10.1096/fj.201801513R

    Figure Lengend Snippet: Effects of IL-1α knockdown. A ) IL-1α expression was knocked down using IL-1α siRNA for 48 h prior to IL-1β stimulation for 24 h. We observed that knocking down IL-1α also reduced IL-1β expression. B ) We reconfirmed the knockdown of IL-1α using an IL-1α ELISA and stimulating RASFs with ATP (5 nM) and LPS (10 μg/ml) to trigger IL-1α release into conditioned medium. C ) Upon confirming IL-1α has been knocked down, we observed a statistically significant decrease of IL-6 production upon IL-1β stimulation. D ) We also observed a decrease in IL-8 production; however, this was not statistically significant. E ) In addition to released proteins, we analyzed expression of cyclooxygenases 1 and 2 (COX-1 and -2) and saw a statistically significant reduction in IL-1β–induced COX-2 and intercellular adhesion molecule 1 expression in IL-1α knockdown samples. * P

    Article Snippet: Small interfering RNA (siRNA) sequences for IL-1α were purchased from MilliporeSigma (eMission predesigned siRNA sequences SASI_Hs01_00167953 and SASI_Hs02_00302829).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Cell fractionation and ChIP findings provide evidence for the role of NF-κBp65 in transcriptional regulation of IL-1α and IL-1β genes. A ) RASFs were stimulated with IL-1β for 24 h prior to collecting, cytoplasmic, nuclear, chromatin bound, and insoluble nuclear fractions which were probed for both mature and pro forms of IL-1β and IL-1α. B ) RASFs lysate (600 µg) from each treated sample was incubated with 30 µl of protein G Sepharose beads along with 3 µg IL-1α pAb. Pull-down proteins were eluted with 2× SDS and loaded onto acrylamide gel for Western blot analysis for interacting partners. C , D ) We performed ChIP assay on the 3 different promoter regions of IL-1α (−418 to −574 upstream of TSS) ( C ) and IL-1β (−64 to −222 upstream of TSS) ( D ) to identify the role of IL-1α and NF-κBp65. NS, nonstimulated.

    Journal: The FASEB Journal

    Article Title: Critical role of IL-1α in IL-1β–induced inflammatory responses: cooperation with NF-κBp65 in transcriptional regulation

    doi: 10.1096/fj.201801513R

    Figure Lengend Snippet: Cell fractionation and ChIP findings provide evidence for the role of NF-κBp65 in transcriptional regulation of IL-1α and IL-1β genes. A ) RASFs were stimulated with IL-1β for 24 h prior to collecting, cytoplasmic, nuclear, chromatin bound, and insoluble nuclear fractions which were probed for both mature and pro forms of IL-1β and IL-1α. B ) RASFs lysate (600 µg) from each treated sample was incubated with 30 µl of protein G Sepharose beads along with 3 µg IL-1α pAb. Pull-down proteins were eluted with 2× SDS and loaded onto acrylamide gel for Western blot analysis for interacting partners. C , D ) We performed ChIP assay on the 3 different promoter regions of IL-1α (−418 to −574 upstream of TSS) ( C ) and IL-1β (−64 to −222 upstream of TSS) ( D ) to identify the role of IL-1α and NF-κBp65. NS, nonstimulated.

    Article Snippet: Small interfering RNA (siRNA) sequences for IL-1α were purchased from MilliporeSigma (eMission predesigned siRNA sequences SASI_Hs01_00167953 and SASI_Hs02_00302829).

    Techniques: Cell Fractionation, Chromatin Immunoprecipitation, Incubation, Acrylamide Gel Assay, Western Blot

    Signaling kinetics of IL-1α and IL-1β corroborate differences in the interaction with IL-1R. A ) RASFs were stimulated with 5 ng/ml of IL-1β or IL-1α for 5–30 min. Key signaling proteins in the IL-1 signaling panel were analyzed using Western immunoblotting. B , C ) Electrostatic potential map and molecular docking studies of the surface of IL-1R protein receptor bound to IL-1α ( B ) and IL-1β ( C ). The electrostatic potential map of interacting surface was made using the last frame after 100 ns molecular dynamics simulation. The interacting surface of the conformation of IL-1R (green) and IL-1α or IL-1β (gray) complex at 100 ns molecular dynamics simulation. The electrostatic potential map of interacting surface of receptor shown in red, blue, and white represent electronegative, electropositive, and hydrophobic surface area of the binding sites, respectively. NS, nonstimulated.

    Journal: The FASEB Journal

    Article Title: Critical role of IL-1α in IL-1β–induced inflammatory responses: cooperation with NF-κBp65 in transcriptional regulation

    doi: 10.1096/fj.201801513R

    Figure Lengend Snippet: Signaling kinetics of IL-1α and IL-1β corroborate differences in the interaction with IL-1R. A ) RASFs were stimulated with 5 ng/ml of IL-1β or IL-1α for 5–30 min. Key signaling proteins in the IL-1 signaling panel were analyzed using Western immunoblotting. B , C ) Electrostatic potential map and molecular docking studies of the surface of IL-1R protein receptor bound to IL-1α ( B ) and IL-1β ( C ). The electrostatic potential map of interacting surface was made using the last frame after 100 ns molecular dynamics simulation. The interacting surface of the conformation of IL-1R (green) and IL-1α or IL-1β (gray) complex at 100 ns molecular dynamics simulation. The electrostatic potential map of interacting surface of receptor shown in red, blue, and white represent electronegative, electropositive, and hydrophobic surface area of the binding sites, respectively. NS, nonstimulated.

    Article Snippet: Small interfering RNA (siRNA) sequences for IL-1α were purchased from MilliporeSigma (eMission predesigned siRNA sequences SASI_Hs01_00167953 and SASI_Hs02_00302829).

    Techniques: Western Blot, Binding Assay

    IL-1β selectively induces IL-1α expression in RASFs. A ) Human samples for RA and normal (NL) synovial tissues (STs) were prepared and analyzed for IL-1α and IL-1β expression. IL-1α and IL-1β expression were significantly higher in RASTs than nondiseased synovial tissues (NLSTs). B ) RASFs were stimulated with IL-1β (10 ng/ml), TNF-α (20 ng/ml), and LPS (5 µg/ml) for 6 h for IL-1α mRNA analysis using quantitative RT-PCR. IL-1β stimulation increases IL-1α mRNA expression significantly. C ) RNA analysis was confirmed using immunofluorescence studies for IL-1α expression. Images were taken using constant exposure was applied across all the samples using Leica immunofluorescence microscope. D ) RASFs were stimulated with varying concentrations of IL-1β (0.5–10 ng/ml) for 24 h then analyzed for IL-1α and IL-1β expression. IL-1α expression increases with as little as 0.5 ng/ml IL-1β. * P

    Journal: The FASEB Journal

    Article Title: Critical role of IL-1α in IL-1β–induced inflammatory responses: cooperation with NF-κBp65 in transcriptional regulation

    doi: 10.1096/fj.201801513R

    Figure Lengend Snippet: IL-1β selectively induces IL-1α expression in RASFs. A ) Human samples for RA and normal (NL) synovial tissues (STs) were prepared and analyzed for IL-1α and IL-1β expression. IL-1α and IL-1β expression were significantly higher in RASTs than nondiseased synovial tissues (NLSTs). B ) RASFs were stimulated with IL-1β (10 ng/ml), TNF-α (20 ng/ml), and LPS (5 µg/ml) for 6 h for IL-1α mRNA analysis using quantitative RT-PCR. IL-1β stimulation increases IL-1α mRNA expression significantly. C ) RNA analysis was confirmed using immunofluorescence studies for IL-1α expression. Images were taken using constant exposure was applied across all the samples using Leica immunofluorescence microscope. D ) RASFs were stimulated with varying concentrations of IL-1β (0.5–10 ng/ml) for 24 h then analyzed for IL-1α and IL-1β expression. IL-1α expression increases with as little as 0.5 ng/ml IL-1β. * P

    Article Snippet: Small interfering RNA (siRNA) sequences for IL-1α were purchased from MilliporeSigma (eMission predesigned siRNA sequences SASI_Hs01_00167953 and SASI_Hs02_00302829).

    Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Microscopy