Proteins detected by Western blot. The presence of the SUMO E2 ligase Ubc9 in the absence (lane 2, left panel) or presence (lane 7, left panel) of co-expressed SUMO-1 did not increase FOG-2 SUMOylation. An inactive mutant of Ubc9 (Ubc9C93S) was used as negative control (lane 3, left panel). Co-expression of FOG-2 with a minimal amount of GFP-SUMO-1 plasmid (100 ng) led to weak FOG-2 SUMOylation (lane 1, right panel, arrowheads). Co-transfection of the indicated SUMO E3 ligases did not 1655472 increase FOG-2 SUMOylation (lanes 2 to 5, right panel). In fact there was a decrease in FOG-2 SUMOylation in the presence of Ubc9 (lane 7, left panel) or E3 ligases (lanes 2 to 5, right panel). This is likely due to the depletion of available SUMO due to the E2- and E3-mediated increase in SUMOylation of other cellular proteins. Together, these experiments indicate that,in COS-7 cells the SUMOylation of FOG-2 is not influenced by co-expression of E2 or E3 ligases. (B) Nuclear localization in HeLa cells. HeLa cells were transfected with GFP-FOG-2 or GFP-FOG2-4KR fusion proteins as indicated in the figure. The cell nuclei were stained with PI (red). There was no detectable difference in the sub-cellular or DprE1-IN-2 web sub-nuclear distribution of wt and mutant FOG-2. Asterisks indicate non-specific bands detected by the FOG-2 antibody. IB, immunoblot. (TIF)AcknowledgmentsWe thank Dr Feng Yan for critical review of the manuscript and helpful discussions.Author ContributionsConceived and designed the experiments: JP BHC. Performed the experiments: JP XMJ DRC. Analyzed the data: JP DC LMK. Wrote the paper: JP BHC.
Primary infection with the human immunodeficiency virus type1 (HIV) is a crucial moment for establishing relationships between virus and host [1,2,3]. The high plasma viral load (pVL) causes a relevant and persistent immune activation that can trigger apoptosis [6?], and becomes chronic in the absence of a valid immune response or without efficient antiretroviral therapy. The immune activation present in this phase is recognizable by typical changes [4], such as an increase in activated/memory CD8+ T cells that express CD38, CD45R0, human leukocyte antigen-DR, and high amounts of cell adhesion molecules, and which canrepresent most part of circulating lymphocytes; a decrease in CD4+ T cells is not always present. High plasma levels of proinflammatory cytokines have been described, along with changes in mitochondrial functionality, augmented tendency to apoptosis and expression of cell death markers (such as CD95) in almost all white blood cells [5,6,7]. However, no gross alterations in Vb T-cell Mirin site repertoire have been found, and the functionality of the T cell repertoire seems well preserved [8]. In turn, immune activation can promote viral replication, so facilitating the infection of other T cells [9,10]. Several studies, including those in animal models, where primary infection has been experimentally induced and strictly monitored, showed that a strictBiomarkers of HIV Control after PHIcorrelation exists between immune activation and progression of the infection 1081537 [11]. During PHI, the appearance of virus-specific cytotoxic T lymphocytes (CTL) coincides with the decay of viral replication, so that patients with a high frequency of HIV-specific CTL display a low pVL and a slow decrease in CD4+ T cell count [12,13]. A significant direct association between the frequency of CD8+ gag-specific T cells and the length of AIDS-free period has been observed during chronic.Proteins detected by Western blot. The presence of the SUMO E2 ligase Ubc9 in the absence (lane 2, left panel) or presence (lane 7, left panel) of co-expressed SUMO-1 did not increase FOG-2 SUMOylation. An inactive mutant of Ubc9 (Ubc9C93S) was used as negative control (lane 3, left panel). Co-expression of FOG-2 with a minimal amount of GFP-SUMO-1 plasmid (100 ng) led to weak FOG-2 SUMOylation (lane 1, right panel, arrowheads). Co-transfection of the indicated SUMO E3 ligases did not 1655472 increase FOG-2 SUMOylation (lanes 2 to 5, right panel). In fact there was a decrease in FOG-2 SUMOylation in the presence of Ubc9 (lane 7, left panel) or E3 ligases (lanes 2 to 5, right panel). This is likely due to the depletion of available SUMO due to the E2- and E3-mediated increase in SUMOylation of other cellular proteins. Together, these experiments indicate that,in COS-7 cells the SUMOylation of FOG-2 is not influenced by co-expression of E2 or E3 ligases. (B) Nuclear localization in HeLa cells. HeLa cells were transfected with GFP-FOG-2 or GFP-FOG2-4KR fusion proteins as indicated in the figure. The cell nuclei were stained with PI (red). There was no detectable difference in the sub-cellular or sub-nuclear distribution of wt and mutant FOG-2. Asterisks indicate non-specific bands detected by the FOG-2 antibody. IB, immunoblot. (TIF)AcknowledgmentsWe thank Dr Feng Yan for critical review of the manuscript and helpful discussions.Author ContributionsConceived and designed the experiments: JP BHC. Performed the experiments: JP XMJ DRC. Analyzed the data: JP DC LMK. Wrote the paper: JP BHC.
Primary infection with the human immunodeficiency virus type1 (HIV) is a crucial moment for establishing relationships between virus and host [1,2,3]. The high plasma viral load (pVL) causes a relevant and persistent immune activation that can trigger apoptosis [6?], and becomes chronic in the absence of a valid immune response or without efficient antiretroviral therapy. The immune activation present in this phase is recognizable by typical changes [4], such as an increase in activated/memory CD8+ T cells that express CD38, CD45R0, human leukocyte antigen-DR, and high amounts of cell adhesion molecules, and which canrepresent most part of circulating lymphocytes; a decrease in CD4+ T cells is not always present. High plasma levels of proinflammatory cytokines have been described, along with changes in mitochondrial functionality, augmented tendency to apoptosis and expression of cell death markers (such as CD95) in almost all white blood cells [5,6,7]. However, no gross alterations in Vb T-cell repertoire have been found, and the functionality of the T cell repertoire seems well preserved [8]. In turn, immune activation can promote viral replication, so facilitating the infection of other T cells [9,10]. Several studies, including those in animal models, where primary infection has been experimentally induced and strictly monitored, showed that a strictBiomarkers of HIV Control after PHIcorrelation exists between immune activation and progression of the infection 1081537 [11]. During PHI, the appearance of virus-specific cytotoxic T lymphocytes (CTL) coincides with the decay of viral replication, so that patients with a high frequency of HIV-specific CTL display a low pVL and a slow decrease in CD4+ T cell count [12,13]. A significant direct association between the frequency of CD8+ gag-specific T cells and the length of AIDS-free period has been observed during chronic.
