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    R&D Systems il 17a
    Systemic immune responses in WT and T-bet −/− mice. Antigen-stimulated splenocyte production of IFN-γ (A) was reduced in T-bet −/− mice; however, there were no differences in IL-4 (B), IL-5 (C), <t>IL-17A</t> (D), or TGF-β1
    Il 17a, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    3t3 l1  (ATCC)
    99
    ATCC 3t3 l1
    Expression of aGPCR during adipogenesis of <t>3T3-L1</t> cells. mRNA levels were determined by qPCR at six time points in the 10-day period of adipocyte differentiation and normalized to the housekeeping gene β actin (Ct = 17.23 ± 0.1). ΔCt values at day 0 are noted in dashed line box. Values of each day were further computed as relative fold change over day 0 expression and expression patterns were constructed. Raw data of qPCR measurements are noted in Supplementary Table 5 . Given is the mean ± SEM ( n = 4 biological replicates).
    3t3 L1, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore tris cl
    Expression of aGPCR during adipogenesis of <t>3T3-L1</t> cells. mRNA levels were determined by qPCR at six time points in the 10-day period of adipocyte differentiation and normalized to the housekeeping gene β actin (Ct = 17.23 ± 0.1). ΔCt values at day 0 are noted in dashed line box. Values of each day were further computed as relative fold change over day 0 expression and expression patterns were constructed. Raw data of qPCR measurements are noted in Supplementary Table 5 . Given is the mean ± SEM ( n = 4 biological replicates).
    Tris Cl, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Cambridge Isotope Laboratories nh4 cl
    Expression of aGPCR during adipogenesis of <t>3T3-L1</t> cells. mRNA levels were determined by qPCR at six time points in the 10-day period of adipocyte differentiation and normalized to the housekeeping gene β actin (Ct = 17.23 ± 0.1). ΔCt values at day 0 are noted in dashed line box. Values of each day were further computed as relative fold change over day 0 expression and expression patterns were constructed. Raw data of qPCR measurements are noted in Supplementary Table 5 . Given is the mean ± SEM ( n = 4 biological replicates).
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    Image Search Results


    Systemic immune responses in WT and T-bet −/− mice. Antigen-stimulated splenocyte production of IFN-γ (A) was reduced in T-bet −/− mice; however, there were no differences in IL-4 (B), IL-5 (C), IL-17A (D), or TGF-β1

    Journal:

    Article Title: T-bet Deficiency Attenuates Renal Injury in Experimental Crescentic Glomerulonephritis

    doi: 10.1681/ASN.2007030392

    Figure Lengend Snippet: Systemic immune responses in WT and T-bet −/− mice. Antigen-stimulated splenocyte production of IFN-γ (A) was reduced in T-bet −/− mice; however, there were no differences in IL-4 (B), IL-5 (C), IL-17A (D), or TGF-β1

    Article Snippet: Concentrations of IL-17A and TGF-β1 were measured by ELISA using an IL-17 DuoSet and TGF-β1 ELISA kit (R & D Systems), respectively, as per the manufacturer's protocol.

    Techniques: Mouse Assay

    Intrarenal mRNA expression of prototypical Th1 (A), Th2 (B), and Th17 (C) cytokines at day 21. Kidney mRNA expression of IFN-γ and IL-4 was unchanged in T-bet −/− mice, but expression of IL-17A was increased. * P

    Journal:

    Article Title: T-bet Deficiency Attenuates Renal Injury in Experimental Crescentic Glomerulonephritis

    doi: 10.1681/ASN.2007030392

    Figure Lengend Snippet: Intrarenal mRNA expression of prototypical Th1 (A), Th2 (B), and Th17 (C) cytokines at day 21. Kidney mRNA expression of IFN-γ and IL-4 was unchanged in T-bet −/− mice, but expression of IL-17A was increased. * P

    Article Snippet: Concentrations of IL-17A and TGF-β1 were measured by ELISA using an IL-17 DuoSet and TGF-β1 ELISA kit (R & D Systems), respectively, as per the manufacturer's protocol.

    Techniques: Expressing, Mouse Assay

    Expression of aGPCR during adipogenesis of 3T3-L1 cells. mRNA levels were determined by qPCR at six time points in the 10-day period of adipocyte differentiation and normalized to the housekeeping gene β actin (Ct = 17.23 ± 0.1). ΔCt values at day 0 are noted in dashed line box. Values of each day were further computed as relative fold change over day 0 expression and expression patterns were constructed. Raw data of qPCR measurements are noted in Supplementary Table 5 . Given is the mean ± SEM ( n = 4 biological replicates).

    Journal: International Journal of Obesity (2005)

    Article Title: The repertoire of Adhesion G protein-coupled receptors in adipocytes and their functional relevance

    doi: 10.1038/s41366-020-0570-2

    Figure Lengend Snippet: Expression of aGPCR during adipogenesis of 3T3-L1 cells. mRNA levels were determined by qPCR at six time points in the 10-day period of adipocyte differentiation and normalized to the housekeeping gene β actin (Ct = 17.23 ± 0.1). ΔCt values at day 0 are noted in dashed line box. Values of each day were further computed as relative fold change over day 0 expression and expression patterns were constructed. Raw data of qPCR measurements are noted in Supplementary Table 5 . Given is the mean ± SEM ( n = 4 biological replicates).

    Article Snippet: 3T3-L1 cell culture, differentiation, and transfection3T3-L1 CL-173™ cells (ATCC, LGC Standards, Wesel, Germany) were cultured and differentiated as previously described [ ].

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Construct

    Activation of endogenous GPR64 in 3T3-L1 cells and primary adipocytes. a Activation of endogenous GPR64 in 3T3-L1 preadipocytes using 0.5 mM agonistic peptide increases intracellular cAMP. Receptor knockdown with siRNA specific for Gpr64 leads to a significantly reduced cAMP accumulation. A scrambled version of the Stachel-peptide (0.5 mM) does not change intracellular cAMP levels. Given is the mean ± SEM of three independent experiments each performed in triplicates (cAMP concentration: Gpr64 siRNA: 3.56 ± 0.65 nM; control siRNA: 2.83 ± 0.33 nM). b Stimulation of mature 3T3-L1 adipocytes with pGPR64 at given concentrations significantly decreases the amount of secreted adiponectin similar to the effect of isoprenaline. As expected, the scrambled peptide does not have an effect on adiponectin secretion. Depicted is the mean ± SEM of five to six independent experiments performed in duplicates. c Glucose uptake was measured in fully differentiated 3T3-L1 cells after incubation with insulin, 0.5 mM pGPR64, 0.5 mM scGPR64, and 1 µM isoprenaline. Insulin induces a significant increase in glucose uptake which is reduced by isoprenaline. The stimulating peptide pGPR64 shows a trend towards reduced insulin-induced glucose uptake, whereas the scrambled peptide does not have an effect. Given is the mean ± SEM of two (isoprenaline) to four (pGPR64, scGPR64) independent experiments. Basal glucose uptake was 2.14 ± 0.34 dpm/mg protein. d Stimulation of GPR64 with the agonistic peptide pGPR64 (0.5 mM) results in a significantly enhanced lipolysis in mature 3T3-L1 and primary adipocytes. As expected, control stimulation of endogenously expressed β adrenergic receptors with isoprenaline increases lipolysis, while the scrambled peptide (at same concentration as the agonistic peptide) does not alter this function. Shown is the mean ± SEM of eight independent experiments performed in triplicates (3T3-L1, basal glycerol release 27.4 ± 2.36 µg/ml) or six independent experiments done in duplicates (primary adipocytes, basal glycerol release 17.4 ± 1.59 µg/ml). Statistical significance was tested using a paired t -test * p

    Journal: International Journal of Obesity (2005)

    Article Title: The repertoire of Adhesion G protein-coupled receptors in adipocytes and their functional relevance

    doi: 10.1038/s41366-020-0570-2

    Figure Lengend Snippet: Activation of endogenous GPR64 in 3T3-L1 cells and primary adipocytes. a Activation of endogenous GPR64 in 3T3-L1 preadipocytes using 0.5 mM agonistic peptide increases intracellular cAMP. Receptor knockdown with siRNA specific for Gpr64 leads to a significantly reduced cAMP accumulation. A scrambled version of the Stachel-peptide (0.5 mM) does not change intracellular cAMP levels. Given is the mean ± SEM of three independent experiments each performed in triplicates (cAMP concentration: Gpr64 siRNA: 3.56 ± 0.65 nM; control siRNA: 2.83 ± 0.33 nM). b Stimulation of mature 3T3-L1 adipocytes with pGPR64 at given concentrations significantly decreases the amount of secreted adiponectin similar to the effect of isoprenaline. As expected, the scrambled peptide does not have an effect on adiponectin secretion. Depicted is the mean ± SEM of five to six independent experiments performed in duplicates. c Glucose uptake was measured in fully differentiated 3T3-L1 cells after incubation with insulin, 0.5 mM pGPR64, 0.5 mM scGPR64, and 1 µM isoprenaline. Insulin induces a significant increase in glucose uptake which is reduced by isoprenaline. The stimulating peptide pGPR64 shows a trend towards reduced insulin-induced glucose uptake, whereas the scrambled peptide does not have an effect. Given is the mean ± SEM of two (isoprenaline) to four (pGPR64, scGPR64) independent experiments. Basal glucose uptake was 2.14 ± 0.34 dpm/mg protein. d Stimulation of GPR64 with the agonistic peptide pGPR64 (0.5 mM) results in a significantly enhanced lipolysis in mature 3T3-L1 and primary adipocytes. As expected, control stimulation of endogenously expressed β adrenergic receptors with isoprenaline increases lipolysis, while the scrambled peptide (at same concentration as the agonistic peptide) does not alter this function. Shown is the mean ± SEM of eight independent experiments performed in triplicates (3T3-L1, basal glycerol release 27.4 ± 2.36 µg/ml) or six independent experiments done in duplicates (primary adipocytes, basal glycerol release 17.4 ± 1.59 µg/ml). Statistical significance was tested using a paired t -test * p

    Article Snippet: 3T3-L1 cell culture, differentiation, and transfection3T3-L1 CL-173™ cells (ATCC, LGC Standards, Wesel, Germany) were cultured and differentiated as previously described [ ].

    Techniques: Activation Assay, Concentration Assay, Incubation

    Effects of aGPCR knockdown on adipogenesis. 3T3-L1 cells were induced to differentiate under transient knockdown of the given aGPCR and compared with control-transfected cells. a , b , c Significant regulation of the adipogenic marker PPARγ under individual knockdown of six receptors was observed during adipogenesis. During the differentiation, we detected three different patterns of PPARγ expression (see Text for details). d Total lipid accumulation was measured by eluted ORO in day 10 adipocytes under receptor-specific transient knockdown and compared with control-transfected cells (Supplementary Table S7 ). e The count of lipid droplets per field of view (0.2664 mm 2 , minimum 5000 droplets counted per experiment) was lowered under knockdown of four receptors compared with control siRNA-transfected cells (Supplementary Table S7 ). f Lipid droplet size was significantly smaller under knockdown of Gpr124 and Gpr126 compared with control siRNA-transfected cells (Supplementary Table S7 ). g , h , i Analysis of lipid droplet size distribution. Size distribution of control is depicted in white bars (min to max). Given is the mean ± SEM ( n > 3 biological replicates). Statistical significance of PPARγ expression, ORO elution and lipid droplet size and count was identified by paired t -test. Lipid droplet size distribution was tested by two-way ANOVA followed by Dunnett’s test for multiple comparisons. * p

    Journal: International Journal of Obesity (2005)

    Article Title: The repertoire of Adhesion G protein-coupled receptors in adipocytes and their functional relevance

    doi: 10.1038/s41366-020-0570-2

    Figure Lengend Snippet: Effects of aGPCR knockdown on adipogenesis. 3T3-L1 cells were induced to differentiate under transient knockdown of the given aGPCR and compared with control-transfected cells. a , b , c Significant regulation of the adipogenic marker PPARγ under individual knockdown of six receptors was observed during adipogenesis. During the differentiation, we detected three different patterns of PPARγ expression (see Text for details). d Total lipid accumulation was measured by eluted ORO in day 10 adipocytes under receptor-specific transient knockdown and compared with control-transfected cells (Supplementary Table S7 ). e The count of lipid droplets per field of view (0.2664 mm 2 , minimum 5000 droplets counted per experiment) was lowered under knockdown of four receptors compared with control siRNA-transfected cells (Supplementary Table S7 ). f Lipid droplet size was significantly smaller under knockdown of Gpr124 and Gpr126 compared with control siRNA-transfected cells (Supplementary Table S7 ). g , h , i Analysis of lipid droplet size distribution. Size distribution of control is depicted in white bars (min to max). Given is the mean ± SEM ( n > 3 biological replicates). Statistical significance of PPARγ expression, ORO elution and lipid droplet size and count was identified by paired t -test. Lipid droplet size distribution was tested by two-way ANOVA followed by Dunnett’s test for multiple comparisons. * p

    Article Snippet: 3T3-L1 cell culture, differentiation, and transfection3T3-L1 CL-173™ cells (ATCC, LGC Standards, Wesel, Germany) were cultured and differentiated as previously described [ ].

    Techniques: Transfection, Marker, Expressing