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lentiviral based shrna constructs plko  (Addgene inc)
96
Addgene inc lentiviral based shrna constructs plko
Fig. 7 EMILIN-2 triggers TLR-4 activation promoting M1 polarization. A qPCR analyses of the expression of TNF-α (M1 marker) relative to GAPDH in TPA-activated THP-1 cells upon IFN-γ/LPS treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. B qPCR analyses of the expression of CD163 (M2 marker) relative to GAPDH in TPA-activated THP-1 cells upon IL-4 treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. C Western blot analyses of pSTAT3 and pNF-κB in TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. D and E Quantification of the results reported in C. F Left: Western blot analyses of TLR-4 immunoprecipitation with the anti-MyD88 antibody using TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2), vehicle (PBS) or LPS as positive control. Whole lysates were loaded as input. Right: quantification of TLR-4 from three different experimental replicates. G qPCR analyses of the expression of TNF-α relative to GAPDH in TPA-activated THP-1 cells w/wo recombinant EMILIN-2 (rE2), and/or the TLR-4 (TAK-242) and NF-κB (JSH-23) inhibitors; n = 3. H Western blot (left) and quantification (right) of the down-regulation of TLR-4 upon <t>lentiviral</t> transduction with two specific <t>shRNA</t> constructs; <t>pLKO</t> served as a shRNA control (shCTRL). I qPCR analyses of TNF-α expression relative to that of GAPDH in TPA-activated THP-1 cells treated or not with recombinant EMILIN-2 (rE2) upon downregulation of TLR-4. Graphs represent the mean ± SD; P values were obtained using the paired Student’s t-test (A, B, D and E) or the One Way Anova Test (F, G, H and I); * P < 0.05, **P < 0.01, ***P < 0.001, n.s.: P > 0.05
Lentiviral Based Shrna Constructs Plko, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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kazuhiro oka unpublished  (Addgene inc)
93
Addgene inc kazuhiro oka unpublished
Fig. 7 EMILIN-2 triggers TLR-4 activation promoting M1 polarization. A qPCR analyses of the expression of TNF-α (M1 marker) relative to GAPDH in TPA-activated THP-1 cells upon IFN-γ/LPS treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. B qPCR analyses of the expression of CD163 (M2 marker) relative to GAPDH in TPA-activated THP-1 cells upon IL-4 treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. C Western blot analyses of pSTAT3 and pNF-κB in TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. D and E Quantification of the results reported in C. F Left: Western blot analyses of TLR-4 immunoprecipitation with the anti-MyD88 antibody using TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2), vehicle (PBS) or LPS as positive control. Whole lysates were loaded as input. Right: quantification of TLR-4 from three different experimental replicates. G qPCR analyses of the expression of TNF-α relative to GAPDH in TPA-activated THP-1 cells w/wo recombinant EMILIN-2 (rE2), and/or the TLR-4 (TAK-242) and NF-κB (JSH-23) inhibitors; n = 3. H Western blot (left) and quantification (right) of the down-regulation of TLR-4 upon <t>lentiviral</t> transduction with two specific <t>shRNA</t> constructs; <t>pLKO</t> served as a shRNA control (shCTRL). I qPCR analyses of TNF-α expression relative to that of GAPDH in TPA-activated THP-1 cells treated or not with recombinant EMILIN-2 (rE2) upon downregulation of TLR-4. Graphs represent the mean ± SD; P values were obtained using the paired Student’s t-test (A, B, D and E) or the One Way Anova Test (F, G, H and I); * P < 0.05, **P < 0.01, ***P < 0.001, n.s.: P > 0.05
Kazuhiro Oka Unpublished, supplied by Addgene inc, 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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paper n a trp53 loxp loxp mice  (Jackson Laboratory)
86
Jackson Laboratory paper n a trp53 loxp loxp mice
Fig. 7 EMILIN-2 triggers TLR-4 activation promoting M1 polarization. A qPCR analyses of the expression of TNF-α (M1 marker) relative to GAPDH in TPA-activated THP-1 cells upon IFN-γ/LPS treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. B qPCR analyses of the expression of CD163 (M2 marker) relative to GAPDH in TPA-activated THP-1 cells upon IL-4 treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. C Western blot analyses of pSTAT3 and pNF-κB in TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. D and E Quantification of the results reported in C. F Left: Western blot analyses of TLR-4 immunoprecipitation with the anti-MyD88 antibody using TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2), vehicle (PBS) or LPS as positive control. Whole lysates were loaded as input. Right: quantification of TLR-4 from three different experimental replicates. G qPCR analyses of the expression of TNF-α relative to GAPDH in TPA-activated THP-1 cells w/wo recombinant EMILIN-2 (rE2), and/or the TLR-4 (TAK-242) and NF-κB (JSH-23) inhibitors; n = 3. H Western blot (left) and quantification (right) of the down-regulation of TLR-4 upon <t>lentiviral</t> transduction with two specific <t>shRNA</t> constructs; <t>pLKO</t> served as a shRNA control (shCTRL). I qPCR analyses of TNF-α expression relative to that of GAPDH in TPA-activated THP-1 cells treated or not with recombinant EMILIN-2 (rE2) upon downregulation of TLR-4. Graphs represent the mean ± SD; P values were obtained using the paired Student’s t-test (A, B, D and E) or the One Way Anova Test (F, G, H and I); * P < 0.05, **P < 0.01, ***P < 0.001, n.s.: P > 0.05
Paper N A Trp53 Loxp Loxp Mice, supplied by Jackson Laboratory, 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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didier trono unpublished  (Addgene inc)
98
Addgene inc didier trono unpublished
Fig. 7 EMILIN-2 triggers TLR-4 activation promoting M1 polarization. A qPCR analyses of the expression of TNF-α (M1 marker) relative to GAPDH in TPA-activated THP-1 cells upon IFN-γ/LPS treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. B qPCR analyses of the expression of CD163 (M2 marker) relative to GAPDH in TPA-activated THP-1 cells upon IL-4 treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. C Western blot analyses of pSTAT3 and pNF-κB in TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. D and E Quantification of the results reported in C. F Left: Western blot analyses of TLR-4 immunoprecipitation with the anti-MyD88 antibody using TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2), vehicle (PBS) or LPS as positive control. Whole lysates were loaded as input. Right: quantification of TLR-4 from three different experimental replicates. G qPCR analyses of the expression of TNF-α relative to GAPDH in TPA-activated THP-1 cells w/wo recombinant EMILIN-2 (rE2), and/or the TLR-4 (TAK-242) and NF-κB (JSH-23) inhibitors; n = 3. H Western blot (left) and quantification (right) of the down-regulation of TLR-4 upon <t>lentiviral</t> transduction with two specific <t>shRNA</t> constructs; <t>pLKO</t> served as a shRNA control (shCTRL). I qPCR analyses of TNF-α expression relative to that of GAPDH in TPA-activated THP-1 cells treated or not with recombinant EMILIN-2 (rE2) upon downregulation of TLR-4. Graphs represent the mean ± SD; P values were obtained using the paired Student’s t-test (A, B, D and E) or the One Way Anova Test (F, G, H and I); * P < 0.05, **P < 0.01, ***P < 0.001, n.s.: P > 0.05
Didier Trono Unpublished, supplied by Addgene inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Fig. 7 EMILIN-2 triggers TLR-4 activation promoting M1 polarization. A qPCR analyses of the expression of TNF-α (M1 marker) relative to GAPDH in TPA-activated THP-1 cells upon IFN-γ/LPS treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. B qPCR analyses of the expression of CD163 (M2 marker) relative to GAPDH in TPA-activated THP-1 cells upon IL-4 treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. C Western blot analyses of pSTAT3 and pNF-κB in TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. D and E Quantification of the results reported in C. F Left: Western blot analyses of TLR-4 immunoprecipitation with the anti-MyD88 antibody using TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2), vehicle (PBS) or LPS as positive control. Whole lysates were loaded as input. Right: quantification of TLR-4 from three different experimental replicates. G qPCR analyses of the expression of TNF-α relative to GAPDH in TPA-activated THP-1 cells w/wo recombinant EMILIN-2 (rE2), and/or the TLR-4 (TAK-242) and NF-κB (JSH-23) inhibitors; n = 3. H Western blot (left) and quantification (right) of the down-regulation of TLR-4 upon lentiviral transduction with two specific shRNA constructs; pLKO served as a shRNA control (shCTRL). I qPCR analyses of TNF-α expression relative to that of GAPDH in TPA-activated THP-1 cells treated or not with recombinant EMILIN-2 (rE2) upon downregulation of TLR-4. Graphs represent the mean ± SD; P values were obtained using the paired Student’s t-test (A, B, D and E) or the One Way Anova Test (F, G, H and I); * P < 0.05, **P < 0.01, ***P < 0.001, n.s.: P > 0.05

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Colorectal cancer development is affected by the ECM molecule EMILIN-2 hinging on macrophage polarization via the TLR-4/MyD88 pathway.

doi: 10.1186/s13046-022-02271-y

Figure Lengend Snippet: Fig. 7 EMILIN-2 triggers TLR-4 activation promoting M1 polarization. A qPCR analyses of the expression of TNF-α (M1 marker) relative to GAPDH in TPA-activated THP-1 cells upon IFN-γ/LPS treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. B qPCR analyses of the expression of CD163 (M2 marker) relative to GAPDH in TPA-activated THP-1 cells upon IL-4 treatment in the presence of recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. C Western blot analyses of pSTAT3 and pNF-κB in TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2) or vehicle (PBS); n = 3. D and E Quantification of the results reported in C. F Left: Western blot analyses of TLR-4 immunoprecipitation with the anti-MyD88 antibody using TPA-activated THP-1 cells challenged with recombinant EMILIN-2 (rE2), vehicle (PBS) or LPS as positive control. Whole lysates were loaded as input. Right: quantification of TLR-4 from three different experimental replicates. G qPCR analyses of the expression of TNF-α relative to GAPDH in TPA-activated THP-1 cells w/wo recombinant EMILIN-2 (rE2), and/or the TLR-4 (TAK-242) and NF-κB (JSH-23) inhibitors; n = 3. H Western blot (left) and quantification (right) of the down-regulation of TLR-4 upon lentiviral transduction with two specific shRNA constructs; pLKO served as a shRNA control (shCTRL). I qPCR analyses of TNF-α expression relative to that of GAPDH in TPA-activated THP-1 cells treated or not with recombinant EMILIN-2 (rE2) upon downregulation of TLR-4. Graphs represent the mean ± SD; P values were obtained using the paired Student’s t-test (A, B, D and E) or the One Way Anova Test (F, G, H and I); * P < 0.05, **P < 0.01, ***P < 0.001, n.s.: P > 0.05

Article Snippet: For lentiviral production, 293FT cells were co-transfected, using a standard calcium phosphate precipitation, with the lentiviral based shRNA constructs (pLKO) and lentiviral vectors pPAX2 and pVSV-G (Addgene, Watertown, USA).

Techniques: Activation Assay, Expressing, Marker, Recombinant, Western Blot, Immunoprecipitation, Positive Control, Transduction, shRNA, Construct, Control