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GARP does not increase FURIN expression or activity, and does not co-immunoprecipitate with FURIN.A. Expression of FURIN mRNA and protein were analyzed by RT-qPCR and WB in the human cells described in Figure 2. B. FURIN activity was measured 24 hours after transfection of 293 cells. Lysates of transfected cells were incubated with a FURIN fluorogenic substrate directly (top panel), or after capture on plastic-coated anti-FURIN antibody (bottom panel), to measure FURIN-like or FURIN specific activity, respectively. Graphs show mean fluorescence intensity at the indicated time (min) after addition of the substrate. The FURIN inhibitor Dec-RVKR-CMK was added to one condition to verify the specificity of the assay. C. Lysates of cells described in Figure 2 were immunoprecipitated with anti-GARP (IP GARP) or anti-FURIN (IP FURIN) antibodies. Immunoprecipitation products or total cell lysates (25% of input used for IPs) were analyzed by WB with anti-GARP, anti-TGF-β or anti-FURIN antibodies, as indicated.
Image collected and cropped by CiteAb under a CC-BY license from the following publication: GARP is regulated by miRNAs and controls latent TGF-β1 production by human regulatory T cells. PLoS One (2013)
GARP does not increase FURIN expression or activity, and does not co-immunoprecipitate with FURIN.A. Expression of FURIN mRNA and protein were analyzed by RT-qPCR and WB in the human cells described in Figure 2. B. FURIN activity was measured 24 hours after transfection of 293 cells. Lysates of transfected cells were incubated with a FURIN fluorogenic substrate directly (top panel), or after capture on plastic-coated anti-FURIN antibody (bottom panel), to measure FURIN-like or FURIN specific activity, respectively. Graphs show mean fluorescence intensity at the indicated time (min) after addition of the substrate. The FURIN inhibitor Dec-RVKR-CMK was added to one condition to verify the specificity of the assay. C. Lysates of cells described in Figure 2 were immunoprecipitated with anti-GARP (IP GARP) or anti-FURIN (IP FURIN) antibodies. Immunoprecipitation products or total cell lysates (25% of input used for IPs) were analyzed by WB with anti-GARP, anti-TGF-β or anti-FURIN antibodies, as indicated.
Image collected and cropped by CiteAb under a CC-BY license from the following publication: GARP is regulated by miRNAs and controls latent TGF-β1 production by human regulatory T cells. PLoS One (2013)
| Regulatory Status |
RUO – Research Use Only |
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