western blot analysis Search Results


western blot analysis  (Cell Signaling Technology Inc)
86
Cell Signaling Technology Inc western blot analysis
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
Western Blot Analysis, 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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ecl western blot analysis system  (Revvity)
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Revvity ecl western blot analysis system
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
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western blot analysis antibody against producer product number primary  (Cell Signaling Technology Inc)
86
Cell Signaling Technology Inc western blot analysis antibody against producer product number primary
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
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western blotting analysis  (CEM Corporation)
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CEM Corporation western blotting analysis
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
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prism statistical analysis of qrt-pcr, western blot, zg thickness and rosette frequency data  (GraphPad Software Inc)
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GraphPad Software Inc prism statistical analysis of qrt-pcr, western blot, zg thickness and rosette frequency data
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
Prism Statistical Analysis Of Qrt Pcr, Western Blot, Zg Thickness And Rosette Frequency Data, supplied by GraphPad Software Inc, 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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enhanced chemiluminescence substrate  (EASY BIO Inc)
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EASY BIO Inc enhanced chemiluminescence substrate
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
Enhanced Chemiluminescence Substrate, supplied by EASY BIO Inc, 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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western blot analysis of cleaved parp protein expression  (CEM Corporation)
90
CEM Corporation western blot analysis of cleaved parp protein expression
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
Western Blot Analysis Of Cleaved Parp Protein Expression, supplied by CEM Corporation, 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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western blot analysis (wb htlv 2.4)  (Genelabs Diagnostics Pte Ltd)
90
Genelabs Diagnostics Pte Ltd western blot analysis (wb htlv 2.4)
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
Western Blot Analysis (Wb Htlv 2.4), supplied by Genelabs Diagnostics Pte Ltd, 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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western blot analysis  (Zhongding Ltd)
90
Zhongding Ltd western blot analysis
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
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western blot analysis using dhfr, phosphorylated prb (ser 780 ), and gapdh antibodies  (CEM Corporation)
90
CEM Corporation western blot analysis using dhfr, phosphorylated prb (ser 780 ), and gapdh antibodies
Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
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western blot analysis  (AGBIO Diagnostics Co Ltd)
90
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Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by <t>Western</t> <t>blot</t> <t>analysis.</t> C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.
Western Blot Analysis, supplied by AGBIO Diagnostics Co Ltd, 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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Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by Western blot analysis. C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.

Journal: American journal of physiology. Lung cellular and molecular physiology

Article Title: Macrophage micro-RNA-155 promotes lipopolysaccharide-induced acute lung injury in mice and rats.

doi: 10.1152/ajplung.00001.2016

Figure Lengend Snippet: Fig. 7. miR-155 targets SOCS-1 to regulate inflammatory response in macrophages. A: wild-type 3=-UTR of mouse SOCS-1 (SOCS-1 3=-UTR) or miR-155 binding deficient mutant 3=-UTR of mouse SOCS-1 (mutSOCS-1 3=-UTR) was separately inserted into the pMIR-REPORT miRNA expression reporter vector. Then these vectors were combined with either miR-155 mimics mmu-miR-155-5p miRNA precursor (Pre-miR-155) or miRNA precursor negative control (Scramble) to be cotransfected into HEK-293T cells. The luciferase activity of the reporter vector was assayed, and the relative activity was compared between groups. n 3 samples per group. *P 0.05 vs. control miRNA and SOCS-1 3=-UTR cotransfected group. B: BMDMs were transfected with SOCS-1 siRNA or control nonsense siRNA. Protein levels of SOCS-1 in cell lysates after transfection were assayed by Western blot analysis. C and D: BMDMs were differentiated from miR-155/ mice or WT control mice, respectively. Then these cells were transfected with either SOCS-1 siRNA or control nonsense siRNA. Immediately after transfection, BMDMs were treated with LPS (100 ng/ml) for 6 h, and mRNA levels of TNF- (C) and IL-6 (D) were assayed by real-time PCR analysis. n 3 samples per group.*P 0.05 vs. BMDMs from WT mice transfected with control siRNA. #P 0.05 vs. BMDMs from WT mice transfected with SOCS-1 siRNA. E: miR-155/ BMDMs transfected with either Pre-miR-155 or miRNA Scramble Control. Then, these cells were transfected with lentivirus expressing SOCS-1 cDNA tagged with a mutated SOCS-1 3=UTR (mutSCOS-1), which was unable to bind miR-155. An empty lentivirus vector (lentiviral vector) was used as a control. At 72 h after transfection, protein levels of SOCS-1 in transfected cells were assayed by Western blot analysis. F and G: 72 h after BMDMs were transfected with indicated miRNAs and lentivirus, BMDMs were stimulated with LPS for 6 h. mRNA levels of TNF- (F), and IL-6 (G) were assayed by real-time PCR analysis. n 3 samples per group. *P 0.05 vs. BMDMs transfected with Pre-miR-155 and lentiviral vector. #P 0.05 vs. BMDMs transfected with Scramble and mutSOCS-1. §P 0.05 vs. BMDMs transfected with Scramble and lentiviral vector.

Article Snippet: The following antibodies were used as primary antibodies for Western blot analysis: anti-p-65 (4764; Cell Signaling Technology, Danvers, MA); anti-p-p65 (3033; Cell Signaling Technology); anti-I B (4812; Cell Signaling Technology); antip-I B (2859; Cell Signaling Technology); anti-JNK (9258; Cell Signaling Technology); anti-p-JNK (4668; Cell Signaling Technology); anti-P38 (9212; Cell Signaling Technology); anti-p-P38 (4511; Cell Signaling Technology); anti-SHIP-1 (ab59338; Abcam); antiBcl-6 (ab86861; Abcam); anti-SOCS-1 (ab9870; Abcam).

Techniques: Binding Assay, Mutagenesis, Expressing, Plasmid Preparation, Negative Control, Luciferase, Activity Assay, Control, Transfection, Western Blot, Real-time Polymerase Chain Reaction