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sclerotium rolfsii  (ATCC)
94
ATCC sclerotium rolfsii
Sclerotium Rolfsii, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pcreb elisa kits  (Cell Signaling Technology Inc)
86
Cell Signaling Technology Inc pcreb elisa kits
Figure 4. A, i, Effect of CRP on intracellular cAMP levels in HMDMs. The cells were incubated with LPS (500 ng/mL for 24 hours) in the absence or presence of CRP (25 g/mL) for 12 hours. The intracellular cAMP levels were measured by <t>ELISA.</t> ANOVA, P0.01; post hoc Tukey test: *P0.001 vs control; **P0.035 vs LPS alone. The results are meanSD of 3 experiments in duplicate. A, ii, Effect of CRP on AC activity in HMDMs. HMDMs on day 7 were pretreated with CRP for 12 hours followed by LPS challenge for 24 hours in serum-free medium. Membrane fractions were used to measure AC activity by the conversion of 32P-ATP to 32P-cAMP. Data are pre- sented as meanSD of 3 experiments in duplicate. The results are expressed as 32P-cAMP counts/mg protein. *P0.05 vs control, **P0.05 vs LPS alone. B, Effect of cAMP agonists on CRP-mediated IL-10 inhibition. Cells were treated with Db-cAMP (50 mol/L) or Br-cAMP (50 mol/L) 1 hour before CRP treatment (12 hours) followed by LPS challenge for 24 hours. IL-10 was measured in superna- tants. The results are meanSD of 3 experiments in duplicate. ANOVA, §P0.01; post hoc Tukey test: P0.001 vs control; *P0.01 vs LPS; **P0.01 vs CRPLPS. C, Effect of CRP on <t>pCREB/CREB</t> levels. Cells were treated with cAMP agonists 30 minutes before cell harvesting following CRPLPS treatment. Cell lysates were prepared as described in Materials and Methods. C, i, Effect of CRP on ratio of pCREB/CREB. Protein (50 g) was used for measurement of CREB and pCREB by ELISA and ratio of pCREB/CREB was cal- culated. ANOVA, P0.05; post hoc Tukey test: §P0.02 vs control; *P0.03 vs LPS alone; **P0.01 vs CRPLPS. The results are meanSD of 3 experiments in duplicate. C, ii, Western blot analysis depicting effect of CRP on pCREB and CREB done in cell lysates as described in Materials and Methods. Lane 1, control; lane 2, LPS; lane 3, CRPLPS; lane 4, CRPLPS Db-cAMP; lane 5, CRPLPSBr-cAMP. §P0.04 vs control, *P0.05 vs LPS alone, **P0.002 vs CRPLPS (n3 experiments for densitometric ratios).
Pcreb Elisa Kits, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cxcr4 target sites  (Addgene inc)
91
Addgene inc cxcr4 target sites
Figure 4. A, i, Effect of CRP on intracellular cAMP levels in HMDMs. The cells were incubated with LPS (500 ng/mL for 24 hours) in the absence or presence of CRP (25 g/mL) for 12 hours. The intracellular cAMP levels were measured by <t>ELISA.</t> ANOVA, P0.01; post hoc Tukey test: *P0.001 vs control; **P0.035 vs LPS alone. The results are meanSD of 3 experiments in duplicate. A, ii, Effect of CRP on AC activity in HMDMs. HMDMs on day 7 were pretreated with CRP for 12 hours followed by LPS challenge for 24 hours in serum-free medium. Membrane fractions were used to measure AC activity by the conversion of 32P-ATP to 32P-cAMP. Data are pre- sented as meanSD of 3 experiments in duplicate. The results are expressed as 32P-cAMP counts/mg protein. *P0.05 vs control, **P0.05 vs LPS alone. B, Effect of cAMP agonists on CRP-mediated IL-10 inhibition. Cells were treated with Db-cAMP (50 mol/L) or Br-cAMP (50 mol/L) 1 hour before CRP treatment (12 hours) followed by LPS challenge for 24 hours. IL-10 was measured in superna- tants. The results are meanSD of 3 experiments in duplicate. ANOVA, §P0.01; post hoc Tukey test: P0.001 vs control; *P0.01 vs LPS; **P0.01 vs CRPLPS. C, Effect of CRP on <t>pCREB/CREB</t> levels. Cells were treated with cAMP agonists 30 minutes before cell harvesting following CRPLPS treatment. Cell lysates were prepared as described in Materials and Methods. C, i, Effect of CRP on ratio of pCREB/CREB. Protein (50 g) was used for measurement of CREB and pCREB by ELISA and ratio of pCREB/CREB was cal- culated. ANOVA, P0.05; post hoc Tukey test: §P0.02 vs control; *P0.03 vs LPS alone; **P0.01 vs CRPLPS. The results are meanSD of 3 experiments in duplicate. C, ii, Western blot analysis depicting effect of CRP on pCREB and CREB done in cell lysates as described in Materials and Methods. Lane 1, control; lane 2, LPS; lane 3, CRPLPS; lane 4, CRPLPS Db-cAMP; lane 5, CRPLPSBr-cAMP. §P0.04 vs control, *P0.05 vs LPS alone, **P0.002 vs CRPLPS (n3 experiments for densitometric ratios).
Cxcr4 Target Sites, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 4. A, i, Effect of CRP on intracellular cAMP levels in HMDMs. The cells were incubated with LPS (500 ng/mL for 24 hours) in the absence or presence of CRP (25 g/mL) for 12 hours. The intracellular cAMP levels were measured by ELISA. ANOVA, P0.01; post hoc Tukey test: *P0.001 vs control; **P0.035 vs LPS alone. The results are meanSD of 3 experiments in duplicate. A, ii, Effect of CRP on AC activity in HMDMs. HMDMs on day 7 were pretreated with CRP for 12 hours followed by LPS challenge for 24 hours in serum-free medium. Membrane fractions were used to measure AC activity by the conversion of 32P-ATP to 32P-cAMP. Data are pre- sented as meanSD of 3 experiments in duplicate. The results are expressed as 32P-cAMP counts/mg protein. *P0.05 vs control, **P0.05 vs LPS alone. B, Effect of cAMP agonists on CRP-mediated IL-10 inhibition. Cells were treated with Db-cAMP (50 mol/L) or Br-cAMP (50 mol/L) 1 hour before CRP treatment (12 hours) followed by LPS challenge for 24 hours. IL-10 was measured in superna- tants. The results are meanSD of 3 experiments in duplicate. ANOVA, §P0.01; post hoc Tukey test: P0.001 vs control; *P0.01 vs LPS; **P0.01 vs CRPLPS. C, Effect of CRP on pCREB/CREB levels. Cells were treated with cAMP agonists 30 minutes before cell harvesting following CRPLPS treatment. Cell lysates were prepared as described in Materials and Methods. C, i, Effect of CRP on ratio of pCREB/CREB. Protein (50 g) was used for measurement of CREB and pCREB by ELISA and ratio of pCREB/CREB was cal- culated. ANOVA, P0.05; post hoc Tukey test: §P0.02 vs control; *P0.03 vs LPS alone; **P0.01 vs CRPLPS. The results are meanSD of 3 experiments in duplicate. C, ii, Western blot analysis depicting effect of CRP on pCREB and CREB done in cell lysates as described in Materials and Methods. Lane 1, control; lane 2, LPS; lane 3, CRPLPS; lane 4, CRPLPS Db-cAMP; lane 5, CRPLPSBr-cAMP. §P0.04 vs control, *P0.05 vs LPS alone, **P0.002 vs CRPLPS (n3 experiments for densitometric ratios).

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: C-Reactive Protein Decreases Interleukin-10 Secretion in Activated Human Monocyte-Derived Macrophages via Inhibition of Cyclic AMP Production

doi: 10.1161/01.atv.0000241572.05292.fb

Figure Lengend Snippet: Figure 4. A, i, Effect of CRP on intracellular cAMP levels in HMDMs. The cells were incubated with LPS (500 ng/mL for 24 hours) in the absence or presence of CRP (25 g/mL) for 12 hours. The intracellular cAMP levels were measured by ELISA. ANOVA, P0.01; post hoc Tukey test: *P0.001 vs control; **P0.035 vs LPS alone. The results are meanSD of 3 experiments in duplicate. A, ii, Effect of CRP on AC activity in HMDMs. HMDMs on day 7 were pretreated with CRP for 12 hours followed by LPS challenge for 24 hours in serum-free medium. Membrane fractions were used to measure AC activity by the conversion of 32P-ATP to 32P-cAMP. Data are pre- sented as meanSD of 3 experiments in duplicate. The results are expressed as 32P-cAMP counts/mg protein. *P0.05 vs control, **P0.05 vs LPS alone. B, Effect of cAMP agonists on CRP-mediated IL-10 inhibition. Cells were treated with Db-cAMP (50 mol/L) or Br-cAMP (50 mol/L) 1 hour before CRP treatment (12 hours) followed by LPS challenge for 24 hours. IL-10 was measured in superna- tants. The results are meanSD of 3 experiments in duplicate. ANOVA, §P0.01; post hoc Tukey test: P0.001 vs control; *P0.01 vs LPS; **P0.01 vs CRPLPS. C, Effect of CRP on pCREB/CREB levels. Cells were treated with cAMP agonists 30 minutes before cell harvesting following CRPLPS treatment. Cell lysates were prepared as described in Materials and Methods. C, i, Effect of CRP on ratio of pCREB/CREB. Protein (50 g) was used for measurement of CREB and pCREB by ELISA and ratio of pCREB/CREB was cal- culated. ANOVA, P0.05; post hoc Tukey test: §P0.02 vs control; *P0.03 vs LPS alone; **P0.01 vs CRPLPS. The results are meanSD of 3 experiments in duplicate. C, ii, Western blot analysis depicting effect of CRP on pCREB and CREB done in cell lysates as described in Materials and Methods. Lane 1, control; lane 2, LPS; lane 3, CRPLPS; lane 4, CRPLPS Db-cAMP; lane 5, CRPLPSBr-cAMP. §P0.04 vs control, *P0.05 vs LPS alone, **P0.002 vs CRPLPS (n3 experiments for densitometric ratios).

Article Snippet: CREB as well as pCREB ELISA kits and antibodies were from Biosource and Cell Signaling Technology respectively.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Control, Activity Assay, Membrane, Inhibition, Cell Harvesting, Western Blot

Figure 5. Effect of transfection in THP-1 with indicated CREB plasmids on CRP- mediated IL-10 inhibition. THP-1 cells were grown in 12-well plates and trans- fected with indicated CREB plasmids as detailed in Materials and Methods. Forty- eight hours after transfection, THP-1 cells were pretreated with CRP (25 g/mL) for 12 hours followed by LPS (500 ng/mL) for 24 hours. A, Superna- tants were used for IL-10 measurement *P0.05 vs control, **P0.05 vs LPS alone, €P0.01 vs control in transfected cells. B, Cells were used for the mea- surement of total and pCREB. The results are expressed as pCREB/CREB ratio. *P0.03 vs control, **P0.045 vs LPS alone, €P0.029 vs control in transfected cells.

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: C-Reactive Protein Decreases Interleukin-10 Secretion in Activated Human Monocyte-Derived Macrophages via Inhibition of Cyclic AMP Production

doi: 10.1161/01.atv.0000241572.05292.fb

Figure Lengend Snippet: Figure 5. Effect of transfection in THP-1 with indicated CREB plasmids on CRP- mediated IL-10 inhibition. THP-1 cells were grown in 12-well plates and trans- fected with indicated CREB plasmids as detailed in Materials and Methods. Forty- eight hours after transfection, THP-1 cells were pretreated with CRP (25 g/mL) for 12 hours followed by LPS (500 ng/mL) for 24 hours. A, Superna- tants were used for IL-10 measurement *P0.05 vs control, **P0.05 vs LPS alone, €P0.01 vs control in transfected cells. B, Cells were used for the mea- surement of total and pCREB. The results are expressed as pCREB/CREB ratio. *P0.03 vs control, **P0.045 vs LPS alone, €P0.029 vs control in transfected cells.

Article Snippet: CREB as well as pCREB ELISA kits and antibodies were from Biosource and Cell Signaling Technology respectively.

Techniques: Transfection, Inhibition, Control