{"database":"bioimages","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Audrey Dumas"],"journal":["The Journal of Cell Biology"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-JCBD-201503124"],"attach_to":["JCB"],"legend":["Primary human macrophages were non-infected, infected with HIV-1YU-2WT or HIV-1YU-2ΔVpr for 8 days. Cells were treated or not (basal) with nocodazole at 10 µM during 1 hour at 37°C. After washing, cells were fixed (time 0) or incubated at 37°C without nocodazole for the indicated times before fixation. Macrophages were then labeled with an anti-p24, followed by Cy2-anti-mouse IgG  and a recombinant anti-tubulin, followed by Cy3-anti-human IgG. Images were acquired under a spinning disk microscope. Z stacks images were acquired.","MDM were nucleofected at day 5 of differentiation to express HA-Vpr or with the HA plasmid as a control. Five hours later, they were fixed and stained , anti-HA antibodies followed by Cy3-anti-rat IgG (left panels), DAPI (second lane) and an anti-p150, followed by Alexa Fluor 488-anti-mouse IgG (middle panels). Stack of images were acquired and deconvoluted. Combined images and a three dimensional reconstitution are shown (right panel).","Primary human macrophages were infected with HIV-1ADA WT or mock infected for 8 days. The cells were incubated for different times with IgG-SRBCs at 37°C. Macrophages were fixed, permeabilized and labeled with AMCA-anti-rabbit IgG to detect the total SRBCs, anti-p24 followed by Cy2-anti-goat IgG, anti-LAMP1 followed by Cy3-anti-mouse IgG, and anti-tubuline followed by Cy5-anti-human IgG. Wide field fluorescent images were acquired.","Primary human macrophages were infected with HIV-1ADA WT or mock infected for 8 days. The cells were incubated for different times with IgG-SRBCs at 37°C. Macrophages were fixed, permeabilized and labeled with AMCA-anti-rabbit IgG to detect the total SRBCs, anti-p24 followed by Cy2-anti-goat IgG, anti-LAMP1 followed by Cy3-anti-mouse IgG, and anti-tubuline followed by Cy5-anti-human IgG. Wide field fluorescent images were acquired, non deconvoluted.","MDM were nucleofected at day 5 of differentiation to express HA-Vpr or with the HA plasmid as a control. Five hours later, they were fixed and stained , anti-HA antibodies followed by Cy3-anti-rat IgG (left panels), DAPI (second lane) and an anti-p150, followed by Alexa Fluor 488-anti-mouse IgG (middle panels). Stack of images were acquired and non deconvoluted. Combined images and a three dimensional reconstitution are shown (right panel).","MDM were nucleofected at day 5 of differentiation to express HA-Vpr or with the HA plasmid as a control, incubated for 60 min with IgG-SRBCs at 37°C, then fixed and permeabilized. They were labeled and analyzed as described in Figure 2 (E,F). Z stacks of wide field fluorescent images were acquired, deconvoluted and a Z projection (Image J) is shown.","Primary human macrophages were infected with HIV-1ADA WT or mock infected for 8 days. The cells were incubated for different times with IgG-SRBCs at 37°C. Macrophages were fixed, permeabilized and labeled with AMCA-anti-rabbit IgG to detect the total SRBCs, anti-p24 followed by Cy2-anti-goat IgG, anti-LAMP1 followed by Cy3-anti-mouse IgG, and anti-tubuline followed by Cy5-anti-human IgG. Wide field fluorescent images were acquired, deconvoluted.","Primary human macrophages were non-infected or infected with HIV-1YU-2WT for 8 days. The cells were then fixed and stained with an anti-p24, followed by Alexa Fluor 488-anti-goat IgG and an anti-MICAL-L1, followed by Cy3-anti-rabbit IgG. Z stacks of wide field fluorescent images were acquired.","(F-K) HeLa cells were transiently transfected to express HA-Vpr or with the HA plasmid as a control, then fixed and the Duolink PLA technology was used with rabbit anti-HA antibodies to detect Vpr combined with mouse mAb anti-EB1 (left panels and I) or mouse anti-p150Glued (middle panels and J), or with mouse anti-HA to detect Vpr combined with rabbit anti-DHC (right panels and K) (H). Negative control was obtained by omitting anti-HA antibody with mouse anti-p150Glued (F) and positive control was with mouse mAb anti-tubulin and rabbit anti-DHC (G).","Primary human macrophages were non-infected (upper panels), infected with HIV-1YU-2WT (middle panels) or HIV-1YU-2ΔVpr (lower panels) for 8 days. The cells were then fixed and stained with an anti-p24, followed by Cy2-anti-goat IgG (not shown) and an anti-p150Glued, followed by Cy3-anti-mouse IgG. Stack of images were acquired.","Primary human macrophages were non-infected or infected with HIV-1YU-2WT for 8 days. The cells were then fixed and stained with an anti-p24, followed by Alexa Fluor 488-anti-goat IgG and an anti-EHD3, followed by Cy3-anti-mouse IgG. Z stacks of wide field fluorescent images were acquired and the stack was deconvoluted.","Primary human macrophages were non-infected or infected with HIV-1Gag-iGFP for 8 days. Gallery of phase contrast images of non-infected (upper panels, see also Movie S1) and HIV-1Gag-iGFP infected (lower panels, see also Movie S2) cells showing phagosome movement.","Primary human macrophages were non-infected or infected with HIV-1ADAWT for 8 days. Total lysates were subjected to western blotting with anti-phospho-ERK1/2 (A), anti-phospho-p38 (B), anti-phospho-SAPK/JNK (C) and anti-phospho-p65/RelA (D). The chemiluminescent signal was quantified and expressed as related to the non-infected condition, showing basal activation by HIV-infection. (E) Macrophages infected for 8 days were incubated for different times with IgG-SRBCs at 37°C, then analyzed by western blot with anti-phospho ERK1/2 and anti-ERK1/2. Results are expressed as a fold increase related to the basal condition for non-infected or HIV-1 infected cells (F). Means ± SEM of three different experiments are plotted. (G) Primary human macrophages were non-infected, infected with HIV-1ADAWT, ","HeLa cells were transiently transfected to express HA-Vpr or with the HA plasmid as a control. Immunoprecipitation with anti-HA antibodies revealed co-immunoprecipiation of endogenous DHC detected with anti-DHC antibodies. The amounts of total proteins in lysates (1% of total lysates) are shown (right panels).","Primary human macrophages were non-infected or infected with HIV-1YU-2WT for 8 days. The cells were then fixed and stained with an anti-p24, followed by Alexa Fluor 488-anti-goat IgG and an anti-EHD3, followed by Cy3-anti-mouse IgG. Z stacks of wide field fluorescent images were acquired.","MDM differentiated for 5 days were treated with control siRNA or siRNA against EB1 for 72h. They were then allowed to phagocytose IgG-opsonized SRBCs for 1h, fixed and stained to detect SRBCs with Alexa Fluor488-anti-rabbit IgG (not shown, red in the merge images, right panels) and LAMP1 with anti-LAMP1 followed by Cy3-anti-mouse IgG (middle panels). SRBCs are also detected with phase contrast (left panels). Z stacks of wide field fluorescent images were acquired, deconvoluted and a Z projection (Image J) is shown","MDM were nucleofected at day 5 of differentiation to express HA-Vpr or with the HA plasmid as a control. Five hours later, they were fixed and stained , anti-HA antibodies followed by Cy3-anti-rat IgG (left panels), DAPI (second lane) and an anti-EB1, followed by Alexa Fluor 488-anti-mouse IgG (middle panels). Stack of images were acquired and non deconvoluted. Combined images and a three dimensional reconstitution are shown (right panel).","MDM were nucleofected at day 5 of differentiation to express HA-Vpr or with the HA plasmid as a control. Five hours later, they were fixed and stained , anti-HA antibodies followed by Cy3-anti-rat IgG (left panels), DAPI (second lane) and an anti-EB1, followed by Alexa Fluor 488-anti-mouse IgG (middle panels). Stack of images were acquired and deconvoluted. Combined images and a three dimensional reconstitution are shown (right panel).","Primary human macrophages were non-infected (upper panels), infected with HIV-1YU-2WT (middle panels) or HIV-1YU-2ΔVpr (lower panels) for 8 days. The cells were then fixed and stained with an anti-p24, followed by Cy2-anti-goat IgG (left panels) and an anti-EB1, followed by Cy3-anti-mouse IgG (middle panels). Stack of images were acquired with a wide field microscope.","Primary macrophages differentiated for 5 days were treated with control siRNA (upper panels) or siRNA against MICAL-L1 (A, lower panels) or siRNA against EHD3 (B, lower panels) for 72h. Cells were then fixed, permeabilized and stained with anti-MICAL-L1 followed by Cy3-F(ab’)2 anti-rabbit IgG (A) or anti-EHD3 followed by Alexa488-F(ab’)2 anti-mouse IgG (B). Stacks of images were acquired on a wide field microscope and a maximum Z projection is shown (left panels). Corresponding phase contrast images are shown (right panels). Bar, 10μm."],"repository":["bioimages"],"figure_sub":["Image 633860 (Figure 3 - C)","Image 633954 (Figure 2 - G)","Figure 8 - C","Figure 8 - A","Image 633966 (Figure 7 - C)","Figure 1 - A-E","Image 633960 (Figure 7 - A)","Image 633996 (Figure 8 - C)","Image 633877 (Figure 6 - A)","Image 633958 (Figure 7 - A)","Image 633874 (Figure 6 - A)","Image 633942 (Figure 2 - G)","Image 633945 (Figure 2 - G)","Image 633969 (Figure 7 - C)","Image 634649 (Figure 1 - B)","Image 633993 (Figure 8 - C)","Image 633976 (Figure 7 - H)","Image 634007 (Supplementary Figure 2 - A)","Image 633889 (Figure 6 - B)","Image 633988 (Figure 8 - A)","Image 633821 (Figure 5 - E)","Image 633974 (Figure 7 - F)","Figure 1 - G","Image 633957 (Figure 2 - G)","Image 633886 (Figure 6 - B)","Image 633883 (Figure 6 - A)","Image 633871 (Figure 6 - A)","Figure 1 - D","Image 634006 (Figure 1 - G)","Figure 1 - C","Image 631193 (Figure 6 - E)","Figure 1 - B","Image 633979 (Figure 7 - H)","Image 633939 (Figure 2 - G)","Image 631194 (Figure 6 - E)","Image 633868 (Figure 6 - A)","Image 631202 (Figure 3 - B)","Image 633863 (Figure 3 - C)","Image 633982 (Figure 8 - A)","Image 634009 (Supplementary Figure 2 - A)","Image 633941 (Figure 2 - G)","Image 633946 (Figure 2 - G)","Image 633992 (Figure 8 - C)","Figure 3 - B","Figure 3 - C","Image 634011 (Supplementary Figure 2 - B)","Image 633882 (Figure 6 - A)","Image 633961 (Figure 7 - A)","Image 633878 (Figure 6 - A)","Image 633984 (Figure 8 - A)","Image 633950 (Figure 2 - G)","Image 633986 (Figure 8 - A)","Image 634651 (Figure 1 - C)","Image 633963 (Figure 7 - A)","Image 633994 (Figure 8 - C)","Image 634008 (Supplementary Figure 2 - A)","Image 634648 (Figure 1 - A-E)","Image 634657 (Figure 7 - E)","Image 633824 (Figure 5 - E)","Image 633971 (Figure 7 - C)","Figure 5 - E","Figure 5","Image 633880 (Figure 6 - A)","Image 633822 (Figure 5 - E)","Figure 7","Image 634655 (Figure 1 - D)","Figure 6","Figure 8","Image 633967 (Figure 7 - C)","Image 633990 (Figure 8 - A)","Image 633876 (Figure 6 - A)","Image 633959 (Figure 7 - A)","Image 634653 (Figure 1 - C)","Image 633948 (Figure 2 - G)","Figure 1","Image 633952 (Figure 2 - G)","Figure 3","Supplementary Figure 2","Figure 2","Image 633980 (Figure 7 - H)","Image 633865 (Figure 3 - C)","Image 633881 (Figure 6 - A)","Image 633965 (Figure 7 - A)","Image 634014 (Supplementary Figure 2 - B)","Figure 7 - A","Figure 7 - C","Image 633938 (Figure 2 - G)","Image 633975 (Figure 7 - G)","Figure 7 - E","Image 633973 (Figure 7 - C)","Figure 7 - F","Figure 7 - G","Image 633983 (Figure 8 - A)","Figure 7 - H","Image 633879 (Figure 6 - A)","Image 633970 (Figure 7 - C)","Image 633962 (Figure 7 - A)","Image 631192 (Figure 6 - E)","Image 631199 (Figure 3 - B)","Image 633859 (Figure 3 - C)","Image 634010 (Supplementary Figure 2 - A)","Image 633867 (Figure 6 - A)","Image 631200 (Figure 3 - B)","Image 633943 (Figure 2 - G)","Image 633944 (Figure 2 - G)","Image 633884 (Figure 6 - A)","Image 633862 (Figure 3 - C)","Image 633890 (Figure 6 - B)","Figure 2 - G","Image 633872 (Figure 6 - A)","Image 633887 (Figure 6 - B)","Image 633956 (Figure 2 - G)","Image 631201 (Figure 3 - B)","Image 633978 (Figure 7 - H)","Image 634013 (Supplementary Figure 2 - B)","Image 633873 (Figure 6 - A)","Image 633997 (Figure 8 - C)","Image 633888 (Figure 6 - B)","Image 633989 (Figure 8 - A)","Image 633864 (Figure 3 - C)","Image 633955 (Figure 2 - G)","Image 633977 (Figure 7 - H)","Image 633866 (Figure 3 - C)","Image 633861 (Figure 3 - C)","Image 633981 (Figure 7 - H)","Image 633885 (Figure 6 - B)","Image 633869 (Figure 6 - A)","Image 633870 (Figure 6 - A)","Image 634647 (Figure 1 - A-E)","Image 633968 (Figure 7 - C)","Image 633823 (Figure 5 - E)","Image 634012 (Supplementary Figure 2 - B)","Image 633949 (Figure 2 - G)","Image 634654 (Figure 1 - D)","Image 633875 (Figure 6 - A)","Image 633985 (Figure 8 - A)","Image 633951 (Figure 2 - G)","Image 633953 (Figure 2 - G)","Image 633940 (Figure 2 - G)","Image 633947 (Figure 2 - G)","Image 633987 (Figure 8 - A)","Image 634650 (Figure 1 - B)","Supplementary Figure 2 - A","Image 633964 (Figure 7 - A)","Image 633991 (Figure 8 - A)","Supplementary Figure 2 - B","Image 633995 (Figure 8 - C)","Image 633972 (Figure 7 - C)","Figure 6 - A","Figure 6 - B","Figure 6 - E"],"pubmed_authors":["Julie Mazzolini","David G. Russell","Serge Benichou","Audrey Dumas","Floriane Herit","Ahmed Zahraoui","Florence Marie-Anaïs","Pierre Bourdoncle","Thomas Guilbert","Florence Niedergang","Gabrielle Lê-Bury"],"additional_accession":[]},"is_claimable":false,"name":"The HIV-1 protein Vpr impairs phagosome maturation by controlling microtubule-dependent trafficking","description":null,"dates":{"release":"2015-10-26T11:27:25Z","modification":"2018-11-29T11:27:25Z","creation":"2018-11-29T11:27:25Z"},"accession":"S-JCBD-201503124","cross_references":{"doi":["10.1083/jcb.201503124"]}}