human il-29 Search Results


il 29 proteins  (Sino Biological)
90
Sino Biological il 29 proteins
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Il 29 Proteins, supplied by Sino Biological, 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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medium by elisa  (R&D Systems)
93
R&D Systems medium by elisa
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Medium By Elisa, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti il 29 antibody  (R&D Systems)
94
R&D Systems anti il 29 antibody
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Anti Il 29 Antibody, supplied by R&D Systems, 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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human il ifnλ  (R&D Systems)
95
R&D Systems human il ifnλ
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Human Il Ifnλ, 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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recombinant human il 29  (R&D Systems)
94
R&D Systems recombinant human il 29
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Recombinant Human Il 29, supplied by R&D Systems, 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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anti ifn λ  (R&D Systems)
92
R&D Systems anti ifn λ
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Anti Ifn λ, supplied by R&D Systems, 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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ifn λ1 il 29 proteins  (Sino Biological)
90
Sino Biological ifn λ1 il 29 proteins
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Ifn λ1 Il 29 Proteins, supplied by Sino Biological, 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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s19 compound pa463  (Chem Impex International)
95
Chem Impex International s19 compound pa463
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
S19 Compound Pa463, supplied by Chem Impex International, 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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human il 29  (R&D Systems)
99
R&D Systems human il 29
RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or <t>IL-29</t> (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.
Human Il 29, supplied by R&D Systems, 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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elisa kits  (R&D Systems)
94
R&D Systems elisa kits
Figure 3. miR-122 <t>enhances</t> <t>IFN</t> response by suppressing p-STAT3. (A) Luciferase activity of a STAT3-responsible promoter construct in HepG2 cells co-transfected with mimics (NC or miR-122) for 2 days, and then transfected with SGR-JFH1 RNA for the indicated time. (B) Western blot analysis of total and phosphorylated STAT3 (p-STAT3, Tyr705) in HepG2 cells first transfected with mimics (NC or miR-122) for 2 days, and then treated with JFH1 RNA or poly(I:C). (C) Analysis of STAT3 protein in HepG2 cells treated with mimics (NC or miR-122) or siRNAs (NC or STAT3). (D, E) Analysis of the mRNAs (D) and proteins (E) of IFNs in HepG2 cells treated with siRNAs (NC or STAT3) and then JFH1 RNA. Cells treated with miR-122 or NC mimics were used as controls in panel D. (F) Analysis of IFN mRNAs in HepG2 cells treated with siRNAs and then with poly(I:C). (G, H) Analysis of p-STAT1 protein and IFN mRNAs in HepG2 cells treated with either S3I-201 or cryptotanshinone (CST) for 24 hr, and then transfected with poly(I:C). Luciferase data are from three experiments (mean +SD). <t>ELISA</t> data are from two experiments (mean +SD). qRT-PCR data are from one experiment that was representative of three experiments (mean ± SEM of technical triplicates). *p < 0.05, **p < 0.01 and ***p < 0.001. DOI: https://doi.org/10.7554/eLife.41159.018 The following source data and figure supplements are available for figure 3:
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tp310515 recombinant human ifn l1 r d systems  (R&D Systems)
93
R&D Systems tp310515 recombinant human ifn l1 r d systems
Figure 3. miR-122 <t>enhances</t> <t>IFN</t> response by suppressing p-STAT3. (A) Luciferase activity of a STAT3-responsible promoter construct in HepG2 cells co-transfected with mimics (NC or miR-122) for 2 days, and then transfected with SGR-JFH1 RNA for the indicated time. (B) Western blot analysis of total and phosphorylated STAT3 (p-STAT3, Tyr705) in HepG2 cells first transfected with mimics (NC or miR-122) for 2 days, and then treated with JFH1 RNA or poly(I:C). (C) Analysis of STAT3 protein in HepG2 cells treated with mimics (NC or miR-122) or siRNAs (NC or STAT3). (D, E) Analysis of the mRNAs (D) and proteins (E) of IFNs in HepG2 cells treated with siRNAs (NC or STAT3) and then JFH1 RNA. Cells treated with miR-122 or NC mimics were used as controls in panel D. (F) Analysis of IFN mRNAs in HepG2 cells treated with siRNAs and then with poly(I:C). (G, H) Analysis of p-STAT1 protein and IFN mRNAs in HepG2 cells treated with either S3I-201 or cryptotanshinone (CST) for 24 hr, and then transfected with poly(I:C). Luciferase data are from three experiments (mean +SD). <t>ELISA</t> data are from two experiments (mean +SD). qRT-PCR data are from one experiment that was representative of three experiments (mean ± SEM of technical triplicates). *p < 0.05, **p < 0.01 and ***p < 0.001. DOI: https://doi.org/10.7554/eLife.41159.018 The following source data and figure supplements are available for figure 3:
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dy7246 primers ifnb1  (R&D Systems)
93
R&D Systems dy7246 primers ifnb1
Figure 3. miR-122 <t>enhances</t> <t>IFN</t> response by suppressing p-STAT3. (A) Luciferase activity of a STAT3-responsible promoter construct in HepG2 cells co-transfected with mimics (NC or miR-122) for 2 days, and then transfected with SGR-JFH1 RNA for the indicated time. (B) Western blot analysis of total and phosphorylated STAT3 (p-STAT3, Tyr705) in HepG2 cells first transfected with mimics (NC or miR-122) for 2 days, and then treated with JFH1 RNA or poly(I:C). (C) Analysis of STAT3 protein in HepG2 cells treated with mimics (NC or miR-122) or siRNAs (NC or STAT3). (D, E) Analysis of the mRNAs (D) and proteins (E) of IFNs in HepG2 cells treated with siRNAs (NC or STAT3) and then JFH1 RNA. Cells treated with miR-122 or NC mimics were used as controls in panel D. (F) Analysis of IFN mRNAs in HepG2 cells treated with siRNAs and then with poly(I:C). (G, H) Analysis of p-STAT1 protein and IFN mRNAs in HepG2 cells treated with either S3I-201 or cryptotanshinone (CST) for 24 hr, and then transfected with poly(I:C). Luciferase data are from three experiments (mean +SD). <t>ELISA</t> data are from two experiments (mean +SD). qRT-PCR data are from one experiment that was representative of three experiments (mean ± SEM of technical triplicates). *p < 0.05, **p < 0.01 and ***p < 0.001. DOI: https://doi.org/10.7554/eLife.41159.018 The following source data and figure supplements are available for figure 3:
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RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or IL-29 (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.

Journal: Journal of Virology

Article Title: Respiratory Syncytial Virus Infection Upregulates NLRC5 and Major Histocompatibility Complex Class I Expression through RIG-I Induction in Airway Epithelial Cells

doi: 10.1128/JVI.00349-15

Figure Lengend Snippet: RSV promotes NLRC5 expression in A549 cells through IFN-β secretion. (A and B) Conditioned medium contains NLRC5-inducing activity. (A) A conditioned medium from RSV-infected A549 cells (CM-RSV A549) was UV irradiated to deactivate RSV (+UV), heat inactivated (HK), or left untreated (−UV). The medium was then mixed with fresh DMEM at a 10% ratio and used to treat A549 cells. (B) Dose response of NLRC5 induction to the conditioned medium. UV-irradiated conditioned medium was mixed with fresh DMEM at various ratios (percent [vol/vol]) and used to treat A549 cells. NLRC5 induction was determined by Western blotting. The expression of viral F protein was determined to show the effect of UV irradiation or heat deactivation on infectious virions. The experiment was performed two times independently. (C) Induction of NLRC5 by recombinant interferons. A549 cells were treated with IFN-β (100 U/ml), IFN-γ (100 U/ml), and IL-28B or IL-29 (10, 30, and 100 ng/ml) for 24 h. Cells were harvested, and NLRC5 induction was detected by Western blotting. GAPDH expression was used as an internal control. (D) Time course of NLRC5 induction by recombinant IFN-β. A549 cells were treated with recombinant IFN-β at 100 U for the indicated times. NLRC5 induction was determined by immunoblotting. GAPDH was the loading control. (E) Immunodepletion of IFN-β removes NLRC5-inducing activity. Aliquots (100 μl) of UV-irradiated conditioned medium were left untreated or incubated at 4°C with anti-IFN-β (10 μg/ml and 30 μg/ml) or anti-IFN-γ (30 μg/ml) for 60 min. The immunocomplexes were removed by protein A-Sepharose beads. The supernatants were mixed with fresh DMEM at 5% (vol/vol) and tested on A549 cells for NLRC5-inducing activity. The results are representative of two independent experiments. GAPDH was used as a loading control. Tx, treatment. (F) Suppression of NLRC5 expression inhibits NLRC5 and MHC-I induction by IFN-β. A549 cells were transfected with scrambled control or with siNLRC5 number 1 (10, 20, or 30 nM) for 24 h. The cells were then treated with 100 U of IFN-β for 24 h. NLRC5 and MHC-I expression was determined by Western blotting. GAPDH expression was the loading control.

Article Snippet: Recombinant human IFN-β, IFN-γ, IL-28B, and IL-29 proteins were purchased from SinoBiologicals Inc. (Beijing, China).

Techniques: Expressing, Activity Assay, Infection, Irradiation, Western Blot, Recombinant, Incubation, Transfection

Figure 3. miR-122 enhances IFN response by suppressing p-STAT3. (A) Luciferase activity of a STAT3-responsible promoter construct in HepG2 cells co-transfected with mimics (NC or miR-122) for 2 days, and then transfected with SGR-JFH1 RNA for the indicated time. (B) Western blot analysis of total and phosphorylated STAT3 (p-STAT3, Tyr705) in HepG2 cells first transfected with mimics (NC or miR-122) for 2 days, and then treated with JFH1 RNA or poly(I:C). (C) Analysis of STAT3 protein in HepG2 cells treated with mimics (NC or miR-122) or siRNAs (NC or STAT3). (D, E) Analysis of the mRNAs (D) and proteins (E) of IFNs in HepG2 cells treated with siRNAs (NC or STAT3) and then JFH1 RNA. Cells treated with miR-122 or NC mimics were used as controls in panel D. (F) Analysis of IFN mRNAs in HepG2 cells treated with siRNAs and then with poly(I:C). (G, H) Analysis of p-STAT1 protein and IFN mRNAs in HepG2 cells treated with either S3I-201 or cryptotanshinone (CST) for 24 hr, and then transfected with poly(I:C). Luciferase data are from three experiments (mean +SD). ELISA data are from two experiments (mean +SD). qRT-PCR data are from one experiment that was representative of three experiments (mean ± SEM of technical triplicates). *p < 0.05, **p < 0.01 and ***p < 0.001. DOI: https://doi.org/10.7554/eLife.41159.018 The following source data and figure supplements are available for figure 3:

Journal: eLife

Article Title: MicroRNA-122 supports robust innate immunity in hepatocytes by targeting the RTKs/STAT3 signaling pathway

doi: 10.7554/elife.41159

Figure Lengend Snippet: Figure 3. miR-122 enhances IFN response by suppressing p-STAT3. (A) Luciferase activity of a STAT3-responsible promoter construct in HepG2 cells co-transfected with mimics (NC or miR-122) for 2 days, and then transfected with SGR-JFH1 RNA for the indicated time. (B) Western blot analysis of total and phosphorylated STAT3 (p-STAT3, Tyr705) in HepG2 cells first transfected with mimics (NC or miR-122) for 2 days, and then treated with JFH1 RNA or poly(I:C). (C) Analysis of STAT3 protein in HepG2 cells treated with mimics (NC or miR-122) or siRNAs (NC or STAT3). (D, E) Analysis of the mRNAs (D) and proteins (E) of IFNs in HepG2 cells treated with siRNAs (NC or STAT3) and then JFH1 RNA. Cells treated with miR-122 or NC mimics were used as controls in panel D. (F) Analysis of IFN mRNAs in HepG2 cells treated with siRNAs and then with poly(I:C). (G, H) Analysis of p-STAT1 protein and IFN mRNAs in HepG2 cells treated with either S3I-201 or cryptotanshinone (CST) for 24 hr, and then transfected with poly(I:C). Luciferase data are from three experiments (mean +SD). ELISA data are from two experiments (mean +SD). qRT-PCR data are from one experiment that was representative of three experiments (mean ± SEM of technical triplicates). *p < 0.05, **p < 0.01 and ***p < 0.001. DOI: https://doi.org/10.7554/eLife.41159.018 The following source data and figure supplements are available for figure 3:

Article Snippet: Recombinant cytokines (IL-29, 1598-IL-025; IFN-b, 8499-IF-010; IL-6, 206-IL-010) and ELISA kits (IL29/IL-28B, DY1598B; IFN-b, DY814) were obtained from R&D systems.

Techniques: Luciferase, Activity Assay, Construct, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR