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erk1/2 mapk inhibitor pd98059  (Merck & Co)
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Merck & Co erk1/2 mapk inhibitor pd98059
A. The distribution of NF-κB p50 and p65, and the levels of p-IκB and IκB in the PAMs inoculated with 1 MOI PCV2 were detected by western blotting at indicated times post-inoculation. B. The levels of Akt, JNK, ERK, p38 MAPK and their phosphorylation in the PAMs inoculated with 1 MOI of PCV2 were detected by western blotting at indicated times post-inoculation. C. The effects of inhibiting NF-κB p50, Akt, ERK and p38 MAPK pathways on the PCV2-induced IL-10. The specific inhibitors of NF-κB (BAY11-7082, 5 μM), PI3K/Akt (LY249002, 10 μM), JNK (SP600125, 10 μM), ERK <t>(PD98059,</t> 20 μM) and p38 MAPK (SB203580, 10 μM) treated cells at 1 h pre-infection, and then cells were infected with 1 MOI of PCV2 for 24 h. The IL-10 secretions were detected by ELISA. D. The efficiency of the siRNAs were evaluated by western blotting after the specific siRNA for NF-κB p50, Akt, p38 and ERK were transfected to cells for 48 h, respectively. E. The effects of downregulating NF-κB p50 (#1), Akt (#1), ERK (#1) and p38 MAPK (#1) on the IL-10 production in PCV2-inoculated PAMs. The most efficiently specific siRNAs or negative control (NC) siRNA transfected PAMs (1×10 6 cells) for 24 h, and PCV2 infected the cells for another 24 h. The IL-10 secretion of each cells were analyzed by ELISA. The data shown are representative of three independent experiments. * P < 0.05, ** P < 0.01 versus DMSO-treated cells (C) or negative control (NC) siRNA-transfected cells (E). # P < 0.05 versus LY-treated cells (C) or Akt specific siRNA-transfected cells (E).
Erk1/2 Mapk Inhibitor Pd98059, supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A. The distribution of NF-κB p50 and p65, and the levels of p-IκB and IκB in the PAMs inoculated with 1 MOI PCV2 were detected by western blotting at indicated times post-inoculation. B. The levels of Akt, JNK, ERK, p38 MAPK and their phosphorylation in the PAMs inoculated with 1 MOI of PCV2 were detected by western blotting at indicated times post-inoculation. C. The effects of inhibiting NF-κB p50, Akt, ERK and p38 MAPK pathways on the PCV2-induced IL-10. The specific inhibitors of NF-κB (BAY11-7082, 5 μM), PI3K/Akt (LY249002, 10 μM), JNK (SP600125, 10 μM), ERK (PD98059, 20 μM) and p38 MAPK (SB203580, 10 μM) treated cells at 1 h pre-infection, and then cells were infected with 1 MOI of PCV2 for 24 h. The IL-10 secretions were detected by ELISA. D. The efficiency of the siRNAs were evaluated by western blotting after the specific siRNA for NF-κB p50, Akt, p38 and ERK were transfected to cells for 48 h, respectively. E. The effects of downregulating NF-κB p50 (#1), Akt (#1), ERK (#1) and p38 MAPK (#1) on the IL-10 production in PCV2-inoculated PAMs. The most efficiently specific siRNAs or negative control (NC) siRNA transfected PAMs (1×10 6 cells) for 24 h, and PCV2 infected the cells for another 24 h. The IL-10 secretion of each cells were analyzed by ELISA. The data shown are representative of three independent experiments. * P < 0.05, ** P < 0.01 versus DMSO-treated cells (C) or negative control (NC) siRNA-transfected cells (E). # P < 0.05 versus LY-treated cells (C) or Akt specific siRNA-transfected cells (E).

Journal: Oncotarget

Article Title: Porcine circovirus type 2 activates PI3K/Akt and p38 MAPK pathways to promote interleukin-10 production in macrophages via Cap interaction of gC1qR

doi: 10.18632/oncotarget.7362

Figure Lengend Snippet: A. The distribution of NF-κB p50 and p65, and the levels of p-IκB and IκB in the PAMs inoculated with 1 MOI PCV2 were detected by western blotting at indicated times post-inoculation. B. The levels of Akt, JNK, ERK, p38 MAPK and their phosphorylation in the PAMs inoculated with 1 MOI of PCV2 were detected by western blotting at indicated times post-inoculation. C. The effects of inhibiting NF-κB p50, Akt, ERK and p38 MAPK pathways on the PCV2-induced IL-10. The specific inhibitors of NF-κB (BAY11-7082, 5 μM), PI3K/Akt (LY249002, 10 μM), JNK (SP600125, 10 μM), ERK (PD98059, 20 μM) and p38 MAPK (SB203580, 10 μM) treated cells at 1 h pre-infection, and then cells were infected with 1 MOI of PCV2 for 24 h. The IL-10 secretions were detected by ELISA. D. The efficiency of the siRNAs were evaluated by western blotting after the specific siRNA for NF-κB p50, Akt, p38 and ERK were transfected to cells for 48 h, respectively. E. The effects of downregulating NF-κB p50 (#1), Akt (#1), ERK (#1) and p38 MAPK (#1) on the IL-10 production in PCV2-inoculated PAMs. The most efficiently specific siRNAs or negative control (NC) siRNA transfected PAMs (1×10 6 cells) for 24 h, and PCV2 infected the cells for another 24 h. The IL-10 secretion of each cells were analyzed by ELISA. The data shown are representative of three independent experiments. * P < 0.05, ** P < 0.01 versus DMSO-treated cells (C) or negative control (NC) siRNA-transfected cells (E). # P < 0.05 versus LY-treated cells (C) or Akt specific siRNA-transfected cells (E).

Article Snippet: For specific inhibitors treated cells, PAM cells were pre-incubated with DMSO, or NF-κB inhibitor BAY11-7082 (Merck), PI3K/Akt inhibitor LY294002 (Merck), ERK1/2 MAPK inhibitor PD98059 (Merck), p38 MAPK inhibitor SB203580 (Merck) and JNK inhibitor SP6000125 (Sigma) for 1 h. For siRNA treated cells, PAM cells were transfected with specific siRNAs ( ) or negative control siRNA using Lipofectamine 2000 for 24 h. Then cells were infected with 1 MOI of PCV2 for 1 h, 12 h or 24 h. Cells were harvested to extract total RNA and proteins.

Techniques: Western Blot, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Negative Control

A. Western blot analysis of the phosphorylation levels of Akt, ERK1/2 and p38 MAPK in wild-type and gC1QR−/− cells inoculated with PCV2 for indicated times. B. Mutated gRNA (gRNA-c1qR-2m) sequences for off-targeting gC1QR locus and corresponding protospacer adjacent motifs (PAM). Red bases: mutations; green bases: PAM. C. Western blotting analysis of gC1QR expression in the cells infected lentivirus containing gRNA-c1qR-2 or gRNA-c1qR-2m as well as wild-type cells. D. Immunoprecipitation analysis of the interaction of Cap with gC1QR in wild-type cells and the cells infected lentivirus containing gRNA-c1qR-2m after PCV2 inoculation E. Western blot analysis of Akt phosphorylation in gRNA-c1qR-2m lentivirus infected cells and wild-type cells after PCV2 inoculation. F. ELISA detection of PCV2-induced IL-10 production in wild-type cells, gRNA-c1qR-2 lentivirus-infected cells and gRNA-c1qR-2m lentivirus-infected cells (1×10 6 cells). The data shown are representative of three independent experiments. The results are mean ± SEM of 3 independent experiments. ** P < 0.01 versus PCV2-inoculated wild-type cells.

Journal: Oncotarget

Article Title: Porcine circovirus type 2 activates PI3K/Akt and p38 MAPK pathways to promote interleukin-10 production in macrophages via Cap interaction of gC1qR

doi: 10.18632/oncotarget.7362

Figure Lengend Snippet: A. Western blot analysis of the phosphorylation levels of Akt, ERK1/2 and p38 MAPK in wild-type and gC1QR−/− cells inoculated with PCV2 for indicated times. B. Mutated gRNA (gRNA-c1qR-2m) sequences for off-targeting gC1QR locus and corresponding protospacer adjacent motifs (PAM). Red bases: mutations; green bases: PAM. C. Western blotting analysis of gC1QR expression in the cells infected lentivirus containing gRNA-c1qR-2 or gRNA-c1qR-2m as well as wild-type cells. D. Immunoprecipitation analysis of the interaction of Cap with gC1QR in wild-type cells and the cells infected lentivirus containing gRNA-c1qR-2m after PCV2 inoculation E. Western blot analysis of Akt phosphorylation in gRNA-c1qR-2m lentivirus infected cells and wild-type cells after PCV2 inoculation. F. ELISA detection of PCV2-induced IL-10 production in wild-type cells, gRNA-c1qR-2 lentivirus-infected cells and gRNA-c1qR-2m lentivirus-infected cells (1×10 6 cells). The data shown are representative of three independent experiments. The results are mean ± SEM of 3 independent experiments. ** P < 0.01 versus PCV2-inoculated wild-type cells.

Article Snippet: For specific inhibitors treated cells, PAM cells were pre-incubated with DMSO, or NF-κB inhibitor BAY11-7082 (Merck), PI3K/Akt inhibitor LY294002 (Merck), ERK1/2 MAPK inhibitor PD98059 (Merck), p38 MAPK inhibitor SB203580 (Merck) and JNK inhibitor SP6000125 (Sigma) for 1 h. For siRNA treated cells, PAM cells were transfected with specific siRNAs ( ) or negative control siRNA using Lipofectamine 2000 for 24 h. Then cells were infected with 1 MOI of PCV2 for 1 h, 12 h or 24 h. Cells were harvested to extract total RNA and proteins.

Techniques: Western Blot, Expressing, Infection, Immunoprecipitation, Enzyme-linked Immunosorbent Assay