{"database":"bioimages","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Fubito Nakatsu"],"journal":["The Journal of Cell Biology"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-JCBD-201206095"],"attach_to":["JCB"],"legend":["Imaging of Golgi complex PtdIns4P  in control MEFs using GFP-GOLPH3.","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), colocalize with the endocytic clathrin machinery component Dynamin 2 (red) in PI4KIII[alpha] KO MEFs.","TIRF microscopic analysis of PI4KIII[alpha] KO MEFs transfected with STIM1-mRFP before the addition of thapsigargin.","F-actin in control MEFs as revealed by staining with fluorescently-labeled phalloidin.","Confocal imaging of live HeLa cells transfected with C6S/C7S/C8S/C9S EFR3A-tdTomato.","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), does not colocalize with the endosomal marker EEA1 (red) in PI4KIII[alpha] KO MEFs.","TIRF microscopic analysis of control MEFs transfected with STIM1-mRFP 8 min after the addition of 1 µm thapsigargin.","Mislocalization of plasma membrane proteins in cells lacking PI4KIII[alpha]. PI4KIII[alpha] KO MEFs were transfected with EFR3B-GFP and imaged by confocal microscopy.","TIRF microscopic analysis of GFP-M1-PI4KIII[alpha] shows transient visits to the plasma membrane (1 s interval between frames).","TIRF microscopic analysis of GFP fluorescence in HeLa cells treated with siRNA against TTC7A/B and transfected with GFP-M1-PI4KIII[alpha].","Chronic depletion of plasma membrane PtdIns(4,5)P2 from HeLa cells by transfection with a membrane-anchored inositol 5-phosphatase (mRFP-IPP1-CAAX, red, see cell on right) results in diffuse intracellular fluorescence of the PtdIns(4,5)P2 probe (iRFP-PH(PLC[delta], blue), but no PtdIns(4,5)P2-positive intracellular vesicles are apparent 16 h post-transfection. Note that the myristoylated/palmitoylated plasma membrane marker L10-GFP (green) is retained at the plasma membrane.","Mislocalization of plasma membrane proteins in cells lacking PI4KIII[alpha]. PI4KIII[alpha] KO MEFs were transfected with M1R-YFP and imaged by confocal microscopy.","Immunofluorescence labeling of endogenous clathrin in control MEFs, as visualized by confocal microscopy.","Imaging analysis of HeLa cells double-transfected with GFP-M1-PI4KIII[alpha] (green) and EFR3B-HA (red) following anti-HA immunfluorescence labeling.","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), does not colocalize with the endosomal marker Rab5 (red) in PI4KIII[alpha] KO MEFs.","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), does not colocalize with the PtdIns3P marker 2xFYVE(Hrs) (red) in PI4KIII[alpha] KO MEFs.","Confocal imaging of live HeLa cells transfected with C5S/C7S/C8S EFR3B-tdTomato.","TIRF microscopic analysis of GFP fluorescence in HeLa cells treated with negative control siRNA and transfected with GFP-M1-PI4KIII[alpha].","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), does not colocalize with the endosomal marker Rab9 (red) in PI4KIII[alpha] KO MEFs.","TIRF microscopic analysis of GFP fluorescence in HeLa cells treated with siRNA against EFR3A/B and TTC7A/B and transfected with GFP-M1-PI4KIII[alpha].","Imaging of Golgi complex PtdIns4P in PI4KIII[alpha] KO MEFs using GFP-GOLPH3.","Palmitoylation of EFR3B is required for recruitment of TTC7B and PI4KIII[alpha] to the plasma membrane. Spinning-disc confocal imaging analysis of HeLa cells transfected with GFP-M1-PI4KIII[alpha] (green), TTC7B-mCherry (red), and the C5S/C7S/C8S triple mutant of EFR3B-FLAG (blue).","TIRF microscopic analysis of PI4KIIIa KO MEFs transfected with STIM1-mRFP 8 min after the addition of 1 µm thapsigargin.","Confocal imaging of live HeLa cells transfected with TTC7B-GFP.","TIRF microscopic analysis of control MEFs transfected with STIM1-mRFP before the addition of thapsigargin.","Immunofluorescence labeling of AP-2 in control MEFs.","Imaging of PtdIns(4,5)P2 distribution in control MEFs using RFP-PH(PLC[delta]).","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), colocalize with the endocytic clathrin machinery component epsin 1 (red) in PI4KIII[alpha] KO MEFs.","Immunofluorescence labeling of AP-2 in PI4KIII[alpha] KO MEFs.","Acute depletion of plasma membrane PtdIns(4,5)P2 using an optogenetic approach results in diffuse intracellular fluorescence of the PtdIns(4,5)P2 probe iRFP-PH(PLC[delta]) but no detectable PtdIns(4,5)P2-positive intracellular vesicles. This image is of iRFP-PH(PLC[delta]) fluorescence before light-mediated PtdIns(4,5)P2 depletion.","TIRF microscopic analysis of PI4KIII[alpha] KO MEFs transfected with STIM1-mRFP and GFP-M1*-PI4KIII[alpha] to rescue before the addition of thapsigargin.","Imaging of PtdIns(4,5)P2 distribution in PI4KIII[alpha] KO MEFs using RFP-PH(PLC[delta]).","Acute depletion of plasma membrane PtdIns(4,5)P2 using an optogenetic approach results in diffuse intracellular fluorescence of the PtdIns(4,5)P2 probe iRFP-PH(PLC[delta]) but no detectable PtdIns(4,5)P2-positive intracellular vesicles. This image is of iRFP-PH(PLC[delta]) fluorescence 1 min after light-mediated PtdIns(4,5)P2 depletion.","Localization of GFP-PH(Osh2) in PI4KIII[alpha] KO MEFs.","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), colocalize with the endocytic clathrin machinery component AP-2 (red) in PI4KIII[alpha] KO MEFs.","TIRF microscopic analysis of GFP fluorescence in HeLa cells treated with siRNA against EFR3A/B and transfected with GFP-M1-PI4KIII[alpha].","Confocal imaging of live HeLa cells transfected with wild-type EFR3A-tdTomato.","Immunofluorescence labeling of endogenous clathrin in PI4KIII[alpha] KO MEFs, as visualized by confocal microscopy.","Imaging of Golgi complex morphology in PI4KIII[alpha] KO MEFs using anti-GM130 immunofluorescence.","Immunofluorescence labeling of epsin 1 in control MEFs.","Imaging analysis of HeLa cells triple-transfected with GFP-M1-PI4KIII[alpha] (green), TTC7B-mCherry (red), and EFR3B-HA (blue). EFR3B-HA immunofluorescence was performed with an anti-HA antibody followed by an Alexa Fluor 647-conjugated anti-rat secondary antibody.","TIRF microscopic analysis of PI4KIII[alpha] KO MEFs transfected with STIM1-mRFP and GFP-M1-PI4KIII[alpha] to rescue before the addition of thapsigargin.","Acute depletion of plasma membrane PtdIns(4,5)P2 using an optogenetic approach results in diffuse intracellular fluorescence of the PtdIns(4,5)P2 probe iRFP-PH(PLC[delta]) but no detectable PtdIns(4,5)P2-positive intracellular vesicles. This image is of iRFP-PH(PLC[delta]) fluorescence 30 min after light-mediated PtdIns(4,5)P2 depletion.","Confocal imaging of HeLa cells transfected with GFP-M1-PI4KIII[alpha].","TIRF microscopic analysis of GFP-M1*-PI4KIII[alpha] shows no transient visits to the plasma membrane (1 s interval between frames).","Colocalization of GFP-PIPK1[gamma] (green) and RFP-PH(PLC[delta]) (red) on intracellular sites in PI4KIII[alpha] KO MEFs as assessed by confocal microscopy.","Acute depletion of plasma membrane PtdIns(4,5)P2 using an optogenetic approach results in diffuse intracellular fluorescence of the PtdIns(4,5)P2 probe iRFP-PH(PLC[delta]) but no detectable PtdIns(4,5)P2-positive intracellular vesicles. This image is of iRFP-PH(PLC[delta]) fluorescence 60 min after light-mediated PtdIns(4,5)P2 depletion.","Confocal imaging of live HeLa cells transfected with EFR3B-tdTomato.","TIRF microscopic analysis of PI4KIII[alpha] KO MEFs transfected with STIM1-mRFP and GFP-M1*-PI4KIII[alpha] to rescue 8 min after the addition of 1 µm thapsigargin.","Imaging analysis of HeLa cells double-transfected with TTC7B-mCherry (red) and EFR3B-HA (green) following anti-HA immunfluorescence labeling.","Imaging of Golgi complex morphology in control MEFs using anti-GM130 immunofluorescence.","TIRF microscopic analysis of GFP fluorescence in HeLa cells treated with negative control siRNA and transfected with GFP-M1*-PI4KIII[alpha].","Localization of GFP-PH(Osh2) in control MEFs.","Immunofluorescence labeling of epsin 1 in PI4KIII[alpha] KO MEFs.","Mislocalization of plasma membrane proteins in cells lacking PI4KIII[alpha]. PI4KIII[alpha] KO MEFs were transfected with L10-GFP and imaged by confocal microscopy.","TIRF microscopic analysis of PI4KIIIa KO MEFs transfected with STIM1-mRFP and GFP-M1-PI4KIIIa to rescue 8 min after the addition of 1 µm thapsigargin.","F-actin in PI4KIII[alpha] KO MEFs as revealed by staining with fluorescently-labeled phalloidin.","Palmitoylation of EFR3B is required for recruitment of TTC7B and PI4KIII[alpha] to the plasma membrane. Spinning-disc confocal imaging analysis of HeLa cells transfected with GFP-M1-PI4KIII[alpha] (green), TTC7B-mCherry (red), and wild-type EFR3B-FLAG (blue).","PtdIns(4,5)P2-positive intracellular vesicles, visualized with PH(PLC[delta]) (green), do not colocalize with the Golgi PtdIns4P marker PH(OSBP) (red) in PI4KIII[alpha] KO MEFs.","Confocal imaging of HeLa cells transfected with GFP-M1*-PI4KIII[alpha]."],"repository":["bioimages"],"figure_sub":["Image 139031 (Figure 7 - G)","Figure 8 - A","Figure 8 - B","Figure 3 - None","Image 139021 (Figure 6 - I)","Image 139005 (Figure 4 - C)","Image 139013 (Figure 6 - A and B)","Image 139280 (Figure 7 - C)","Supplemental Figure 2 - A","Supplemental Figure 2 - B","Image 139398 (Figure 1 - E)","Image 139007 (Figure 5 - G)","Image 138986 (Figure 2 - C)","Image 139056 (Supplemental Figure 5 - A)","Image 139291 (Supplemental Figure 2 - B)","Figure 1 - A","Image 139284 (Figure 8 - A)","Figure 1 - E","Image 139011 (Figure 5 - D)","Image 139285 (Figure 8 - A)","Figure 6 - A and B","Image 139023 (Figure 6 - J)","Image 139027 (Figure 7 - A)","Image 139001 (Figure 3 - None)","Image 138999 (Figure 3 - None)","Image 139032 (Figure 7 - G)","Image 139052 (Figure 8 - B)","Image 139010 (Figure 5 - D)","Image 139003 (Figure 4 - C)","Image 138964 (Figure 1 - A)","Figure 6 - G and H","Figure 5 - F","Image 139008 (Figure 5 - F)","Figure 5 - G","Image 139054 (Figure 8 - B)","Image 139399 (Figure 1 - E)","Figure 5 - D","Image 139288 (Figure 8 - A)","Figure 5","Figure 4","Figure 7","Image 139293 (Figure 2 - E)","Figure 6","Figure 8","Figure 1","Supplemental Figure 5 - A","Figure 3","Figure 2","Supplemental Figure 5","Supplemental Figure 2","Figure 7 - A","Figure 7 - C","Image 139286 (Figure 8 - A)","Figure 7 - G","Figure 7 - H","Figure 6 - C and D","Image 139025 (Figure 6 - K)","Image 139009 (Figure 5 - F)","Image 139283 (Figure 8 - A)","Image 138987 (Figure 2 - C)","Image 138998 (Figure 3 - None)","Image 139290 (Supplemental Figure 2 - A)","Figure 2 - E","Figure 2 - F","Image 139006 (Figure 5 - G)","Image 139030 (Figure 7 - G)","Figure 2 - C","Figure 2 - D","Image 139014 (Figure 6 - C and D)","Image 139055 (Supplemental Figure 5 - A)","Image 139282 (Figure 8 - A)","Image 139017 (Figure 6 - E and F)","Figure 6 - E and F","Image 139019 (Figure 6 - G and H)","Image 139002 (Figure 3 - None)","Figure 4 - C","Image 139287 (Figure 8 - A)","Figure 6 - K","Image 139018 (Figure 6 - G and H)","Image 139033 (Figure 7 - G)","Image 139015 (Figure 6 - C and D)","Figure 6 - I","Image 139294 (Figure 2 - F)","Figure 6 - J","Image 139012 (Figure 6 - A and B)","Image 139053 (Figure 8 - B)","Image 139024 (Figure 6 - K)","Image 138965 (Figure 1 - A)","Image 139020 (Figure 6 - I)","Image 139016 (Figure 6 - E and F)","Image 139000 (Figure 3 - None)","Image 139022 (Figure 6 - J)","Image 139026 (Figure 7 - A)","Image 139292 (Figure 2 - D)","Image 139004 (Figure 4 - C)","Image 139289 (Figure 8 - A)","Image 139281 (Figure 7 - H)"],"pubmed_authors":["Lukas B. Tanner","Mingming Hao","Markus R. Wenk","Fubito Nakatsu","Guanghou Shui","Michelle Pirruccello","Sang Yoon Lee","Nicholas T. Ingolia","Jeeyun Chung","Pietro De Camilli","Jeremy M. Baskin"],"additional_accession":[]},"is_claimable":false,"name":"PtdIns4P synthesis by PI4KIIIα at the plasma membrane and its impact on plasma membrane identity","description":null,"dates":{"release":"2012-12-10T11:23:39Z","modification":"2018-11-29T11:23:39Z","creation":"2018-11-29T11:23:39Z"},"accession":"S-JCBD-201206095","cross_references":{"doi":["10.1083/jcb.201206095"]}}