{"database":"bioimages","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Eva Nievergall"],"journal":["The Journal of Cell Biology"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-JCBD-201005035"],"attach_to":["JCB"],"legend":["EphrinA5-Fc stimulated (10min) EphA3/HEK293T cells expressing exogenous w/t GFP-PTP1B were fixed, permeabilised and labelled with anti-EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. Alexa 546 fluorescent channel is shown.","EphA3/HEK293T cells expressing exogenous GFP-PTP1B-[D-A], whithout ephrin-A5 stimulation, were fixed, permeabilised and labelled with anti-EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. GFP fluorescent channel is shown.","EphrinA5-Fc stimulated (10min) EphA3/HEK293T cells expressing exogenous w/t GFP-PTP1B were fixed, permeabilised and labelled with anti-EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. GFP fluorescent channel is shown.","EphA3/HEK293T cells expressing exogenous GFP-PTP1B-[D-A] or w/t GFP-PTP1B, without EphrinA5-Fc stimulation, were fixed, permeabilised and labelled with -EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. GFP channel is shown.","EphA3/HEK293T cells expressing exogenous GFP-PTP1B-[D-A] or w/t GFP-PTP1B, without EphrinA5-Fc stimulation, were fixed, permeabilised and labelled with -EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. Alexa546 fluorescent channel is shown.","Cell segregation was monitored in co-cultures of U251 cells and cell tracker green-labelled HEK293 cells in the presence or absence (control) of PTP1B inhibitor (PTP1B-I, 10µM), EphA3 inhibitor (EphA3-I, 10µM) or DMSO (as control for EphA3-I) for 72h. Bright-field, fluorescence and merged images of 4 separate PFA-fixed cell cultures are shown","EphA3/HEK293T cells expressing exogenous GFP-PTP1B-[D-A], whithout ephrin-A5 stimulation, were fixed, permeabilised and labelled with anti-EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. Alexa 546 fluorescent channel is shown.","U251 cells treated for 48 hours with PTP1B-targeted shRNA lentivirus, as indicated, were plated with HEK293 cells, using cell tracker green to differentiate populations. Upon completion of segregation, fixed cells were analysed by fluorescence microscopy and image analysis. Low magnification (4x) images analysed in are shown and used for analysis by estimating and plotting the overall intensity per area of cell tracker green fluorescence (n = 4).","COS7 cells, co-transfected with GFP-PTP1B-[D-A] and EphA3 were stimulated with Alexa594-ephrinA5-Fc-coated Protein A-Dynabeads to cluster EphA3 and triggering recruitment of ER-resident PTP1B to the activated receptor. In this setting, shedding of ephrin and internalisation of EphA3/ephrin complexes results in the appearance of punctuate clusters around the dynabeads. Fixed cells were analysed by confocal microscopy.","EphrinA5-Fc stimulated (10min) EphA3/HEK293T cells expressing exogenous GFP-PTP1B-[D-A] were fixed, permeabilised and labelled with -EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. Alexa546 fluorescent channel is shown.","COS7 cells, transfected with GFP-PTP1B-[D-A] and APN-EphA3, when co-transfected with TM-BirA, yielded biotinylated EphA3 that can be stimulated with SA-Dynabeads. Cell surface EphA3 was labelled on intact  with anti-EphA3 mAb and detected with Alexa546-labelled secondary antibodies by confocal microscopy. SA Dynabeads are shown were PTP1B/EphA3 co-localisation is apparent.COS7 cells, transfected with GFP-PTP1B-[D-A] and APN-EphA3, when co-transfected with TM-BirA, yielded biotinylated EphA3 that can be stimulated with SA-Dynabeads. Cell surface EphA3 was labelled on intact cells with IIIA4","EphrinA5-Fc stimulated (10min) EphA3/HEK293T cells expressing exogenous GFP-PTP1B-[D-A] were fixed, permeabilised and labelled with -EphA3 and Alexa546-labelled secondary antibodies for confocal microscopy. GFP fluorescent channel is shown.","U251 cells treated for 48 hours with non-targeted shRNA lentivirus, were plated with HEK293 cells, using cell tracker green to differentiate populations. Upon completion of segregation, fixed cells were analysed by fluorescence microscopy and image analysis. Low magnification (4x) images analysed in are shown and used for analysis by estimating and plotting the overall intensity per area of cell tracker green fluorescence (n = 4).","COS7 cells, transfected with GFP-PTP1B-[D-A] and APN-EphA3, only when co-transfected with TM-BirA, yielded biotinylated EphA3 that can be stimulated with SA-Dynabeads. Cell surface EphA3 was labelled on intact cells with IIIA4 -EphA3 mAb and detected with Alexa546-labelled secondary antibodies by confocal microscopy. No PTP1B/EphA3 co-localisation with SA Dynabeads is apparent.","Cell segregation was monitored in co-cultures of U251 cells and cell tracker green-labelled HEK293 cells in the presence or absence (control) of PTP1B inhibitor (PTP1B-I, 10µM), EphA3 inhibitor (EphA3-I, 10µM) or DMSO (as control for EphA3-I) for 72h. Bright-field, fluorescence and merged images of 4 separate PFA-fixed cell cultures are shown.","COS7 cells, transfected with GFP-PTP1B-[D-A] and APN-EphA3, when co-transfected with TM-BirA, yielded biotinylated EphA3 that can be stimulated with SA-Dynabeads. Cell surface EphA3 was labelled on intact cells with IIIA4 -EphA3 mAb and detected with Alexa546-labelled secondary antibodies by confocal microscopy. SA Dynabeads are shown were PTP1B/EphA3 co-localisation is apparent.","U251 cells treated for 48 hours with non-targeted or PTP1B-targeted shRNA lentivirus, as indicated, were plated with HEK293 cells, using cell tracker green to differentiate populations. Upon completion of segregation, fixed cells were analysed by fluorescence microscopy and image analysis as outlined in Figure S3. 10x magnification images are shown and used for analysis by quantitation of fluorescence intensity contour plots."],"repository":["bioimages"],"figure_sub":["Image 28554 (Figure 5 - B)","Supplemental Figure 4","Image 28579 (Figure 7 - B)","Image 28604 (Supplemental Figure 4 - A)","Image 28595 (Figure 7 - B)","Figure 8 - A","Image 28491 (Figure 5 - C)","Image 28521 (Supplemental Figure 4 - A)","Image 28558 (Figure 7 - B)","Figure 7 - B","Image 28560 (Figure 7 - B)","Image 28588 (Figure 7 - B)","Image 28590 (Figure 7 - B)","Image 28609 (Supplemental Figure 4 - A)","Image 28510 (Figure 8 - A)","Image 28562 (Figure 7 - B)","Image 28586 (Figure 7 - B)","Image 28556 (Figure 7 - B)","Image 28583 (Figure 7 - B)","Image 28524 (Supplemental Figure 4 - A)","Image 28600 (Figure 7 - B)","Image 28565 (Figure 7 - B)","Image 28610 (Figure 5 - B)","Image 28509 (Figure 8 - A)","Image 28513 (Figure 8 - A)","Image 28606 (Supplemental Figure 4 - A)","Image 28574 (Figure 7 - B)","Image 28603 (Figure 7 - B)","Image 28598 (Figure 7 - B)","Image 28516 (Figure 8 - A)","Image 28523 (Supplemental Figure 4 - A)","Image 28568 (Figure 7 - B)","Image 28584 (Figure 7 - B)","Image 28571 (Figure 7 - B)","Image 28549 (Figure 5 - B)","Image 28607 (Supplemental Figure 4 - A)","Image 28551 (Figure 5 - B)","Image 28553 (Figure 5 - B)","Image 28602 (Figure 7 - B)","Image 28515 (Figure 8 - A)","Image 28526 (Supplemental Figure 4 - A)","Image 28567 (Figure 7 - B)","Image 28597 (Figure 7 - B)","Image 28572 (Figure 7 - B)","Image 28585 (Figure 7 - B)","Image 28552 (Figure 5 - B)","Image 28587 (Figure 7 - B)","Image 28525 (Supplemental Figure 4 - A)","Image 28514 (Figure 8 - A)","Image 28566 (Figure 7 - B)","Image 28582 (Figure 7 - B)","Image 28555 (Figure 5 - B)","Image 28573 (Figure 7 - B)","Image 28601 (Figure 7 - B)","Image 28575 (Figure 7 - B)","Image 28599 (Figure 7 - B)","Image 28570 (Figure 7 - B)","Image 28569 (Figure 7 - B)","Image 28492 (Figure 5 - C)","Image 28596 (Figure 7 - B)","Image 28591 (Figure 7 - B)","Image 28520 (Supplemental Figure 4 - A)","Image 28557 (Figure 7 - B)","Image 28578 (Figure 7 - B)","Image 28561 (Figure 7 - B)","Image 28550 (Figure 5 - B)","Image 28593 (Figure 7 - B)","Image 28563 (Figure 7 - B)","Image 28580 (Figure 7 - B)","Image 28577 (Figure 7 - B)","Image 28512 (Figure 8 - A)","Image 28564 (Figure 7 - B)","Image 28592 (Figure 7 - B)","Figure 5 - B","Figure 5 - C","Figure 5","Image 28519 (Supplemental Figure 4 - A)","Image 28594 (Figure 7 - B)","Figure 7","Supplemental Figure 4 - A","Image 28605 (Supplemental Figure 4 - A)","Figure 8","Image 28581 (Figure 7 - B)","Image 28522 (Supplemental Figure 4 - A)","Image 28490 (Figure 5 - C)","Image 28559 (Figure 7 - B)","Image 28576 (Figure 7 - B)","Image 28589 (Figure 7 - B)","Image 28511 (Figure 8 - A)","Image 28608 (Supplemental Figure 4 - A)"],"pubmed_authors":["Fawaz G. Haj","Benjamin G. Neel","Martin Lackmann","Mary E. Vail","Eva Nievergall","Carolin Stegmayer","Peter W. Janes","Shyh Wei Teng","Philippe I. Bastiaens"],"additional_accession":[]},"is_claimable":false,"name":"PTP1B regulates Eph receptor function and trafficking","description":null,"dates":{"release":"2010-12-06T11:20:30Z","modification":"2018-11-29T11:20:30Z","creation":"2018-11-29T11:20:30Z"},"accession":"S-JCBD-201005035","cross_references":{"doi":["10.1083/jcb.201005035"]}}