ergic-53 Search Results


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  • 93
    Millipore ergic 53
    PLN arginine mutants are mislocalized out of the ER. ( A ) HEK cells were transiently transfected with tagged WT-PLN or PLN mutants and visualized by confocal microscopy to detect the flag tag. Note that mutation of either Arg13 or Arg14, or both together, resulted in mislocalization of PLN out of the ER with significant expression at the plasma membrane. Cells shown represent at least 20 representative cells per condition. ( B ) Primary mouse fibroblasts were transiently transfected with tagged WT-PLN or the RΔ14 PLN construct and visualized by confocal microscopy. Endogenous ER markers <t>ERGIC-53</t> and calreticulin as well as plasma membrane protein markers Na/K ATPase and Na/Ca exchanger (NCX) were probed against to allow accurate visualization of the ER and the plasma membrane. WT-PLN shows staining similar to the ER markers and RΔ14 PLN shows more diffuse ER staining as well as pronounced plasma membrane staining. Inserts shows a higher magnification of the cell membrane. Note, no plasma membrane staining is seen in WT-PLN and the other ER proteins but clear membrane staining is seen for RΔ14 PLN and the plasma membrane proteins NCX and Na/K ATPase. (Scale bar, 10 µm)
    Ergic 53, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology ergic 53
    TANGO1 amino acids 1255–1295 are the minimal TEER. ( A ) A schematic depiction of myc-epitope tagged mitochondrially-targeted (mit-TEER) truncates. ( B ) mit-TEER truncates were expressed in 2H5 cells, fixed and stained with anti-myc-antibody and visualised with confocal microscopy. ( C ) mit-TEER truncates were expressed in 2H5 cells, which were fixed and stained using anti-myc antibody (green) and, as a mitochondrial marker, anti-HSP60 antibody (red). ( D ) Overlap of the signal from myc and HSP60 was quantified and plotted as the Manders’ overlap coefficient for the two constructs (mit-Δ1255–1295 and mit-Δ1296–1336 respectively). ( E ) 2H5 cells were transfected with mit-Δ1255–1295 or mit-Δ1296–1336, fixed, and stained with anti-myc, anti-HSP60 and <t>anti-ERGIC-53</t> antibodies. Arrows indicate myc staining with or without colocalised ERGIC-53 staining. ( F ) The extent of overlap of ERGIC-53 and myc was quantified and plotted as the Manders’ overlap coefficient for mit-Δ1255–1295 and mit-Δ1296–1336, respectively. Scale bars: ( B, C, E and F ) 20 μm; inset 2 μm.
    Ergic 53, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enzo Biochem anti ergic 53
    F805C selectively co-localizes with BAP31. (A) Representative confocal images of HEK293 cells expressing F805C-Kv11.1 protein showing the overlap between anti-Kv11.1 and anti-ER/ERGIC protein immunostaining. Anti-Kv11.1 immunostaining is shown in red; anti-calnexin ( n = 12 images), anti-Sec31 ( n = 19 images), <t>anti-ERGIC-53</t> ( n = 19 images), anti-derlin-1 ( n = 18 images), anti-KDEL ( n = 15 images), or anti-BAP31 ( n = 17 images) is shown as green; overlapping immunostaining of red and green signals of similar intensities is shown as yellow; and the cell nuclei are labeled blue. The arrowheads highlight the distinct immunostaining pattern of F805C-Kv11.1 protein, and the scale bar represents 10 μm. (B) The mean Costes’ Pearson Coefficient (PC) for the image data sets are shown. PC values at or below the dashed line are (PC = 0.5) indicative of weak or no co-localization. Two independent cultures were tested for each condition.
    Anti Ergic 53, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 88/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Axxora ergic 53 antibody
    F805C selectively co-localizes with BAP31. (A) Representative confocal images of HEK293 cells expressing F805C-Kv11.1 protein showing the overlap between anti-Kv11.1 and anti-ER/ERGIC protein immunostaining. Anti-Kv11.1 immunostaining is shown in red; anti-calnexin ( n = 12 images), anti-Sec31 ( n = 19 images), <t>anti-ERGIC-53</t> ( n = 19 images), anti-derlin-1 ( n = 18 images), anti-KDEL ( n = 15 images), or anti-BAP31 ( n = 17 images) is shown as green; overlapping immunostaining of red and green signals of similar intensities is shown as yellow; and the cell nuclei are labeled blue. The arrowheads highlight the distinct immunostaining pattern of F805C-Kv11.1 protein, and the scale bar represents 10 μm. (B) The mean Costes’ Pearson Coefficient (PC) for the image data sets are shown. PC values at or below the dashed line are (PC = 0.5) indicative of weak or no co-localization. Two independent cultures were tested for each condition.
    Ergic 53 Antibody, supplied by Axxora, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore monoclonal anti ergic 53
    Analysis of nocodazole induced Golgi fragmentation in mSar1p injected cells. Vero cells were incubated for 30 min at 4°C to depolymerize microtubules. Cells were microinjected in the cold with 2 mg/ml purified mSar1p mixed with 2 mg/ml cascade blue conjugated BSA. Immediately after injection, cells were incubated for 1 h ( a and b ) or 2 h ( c and d ) at 37°C in the presence of nocodazole (10 μM) before they were fixed and stained for <t>ERGIC</t> 53 ( a , c , and e ) or the Golgi marker giantin ( b , d , and f ). In e and f , cells were first treated with nocodazole for 2 h, microinjected and subsequently incubated in the presence of nocodazole for 1 h. Microinjected cells (marked by asterisks) were identified by the coinjected BSA (not shown). Microinjection of mSar1p results in the accumulation of ERGIC 53 in the ER but does not interfere with the formation and distribution of the Golgi fragments (marked by arrowheads in b ); in noninjected cells ERGIC 53 and giantin often appear to colocalize in peripheral Golgi fragments (marked by arrows). Bar, 15 μm.
    Monoclonal Anti Ergic 53, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Becton Dickinson ergic 53
    Phosphorylation of Sec16 is linked to ERES number. (A–C) Hells cells were transfected with cDNA encoding wild-type GFP-Sec16 (GFP–Sec16-wt; A) or GFP–Sec16-T415I (B), fixed 24 h later, and immunostained for Sec31 to label ERESs or <t>ERGIC-53</t> to label the ERGIC. (C) The number of ERESs and peripheral ERGIC punctae was counted. The bar graph represents the number of punctae positive for Sec31 (ERES) or ERGIC-53 (ERGIC). The values determined in cells expressing GFP–Sec16-wt were set to 100% (means ± SD; three independent experiments; asterisks indicate statistically significant difference to GFP–Sec16-wt; *, P
    Ergic 53, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 89/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Axxora ergic 53
    Phosphorylation of Sec16 is linked to ERES number. (A–C) Hells cells were transfected with cDNA encoding wild-type GFP-Sec16 (GFP–Sec16-wt; A) or GFP–Sec16-T415I (B), fixed 24 h later, and immunostained for Sec31 to label ERESs or <t>ERGIC-53</t> to label the ERGIC. (C) The number of ERESs and peripheral ERGIC punctae was counted. The bar graph represents the number of punctae positive for Sec31 (ERES) or ERGIC-53 (ERGIC). The values determined in cells expressing GFP–Sec16-wt were set to 100% (means ± SD; three independent experiments; asterisks indicate statistically significant difference to GFP–Sec16-wt; *, P
    Ergic 53, supplied by Axxora, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher ergic 53
    Phosphorylation of Sec16 is linked to ERES number. (A–C) Hells cells were transfected with cDNA encoding wild-type GFP-Sec16 (GFP–Sec16-wt; A) or GFP–Sec16-T415I (B), fixed 24 h later, and immunostained for Sec31 to label ERESs or <t>ERGIC-53</t> to label the ERGIC. (C) The number of ERESs and peripheral ERGIC punctae was counted. The bar graph represents the number of punctae positive for Sec31 (ERES) or ERGIC-53 (ERGIC). The values determined in cells expressing GFP–Sec16-wt were set to 100% (means ± SD; three independent experiments; asterisks indicate statistically significant difference to GFP–Sec16-wt; *, P
    Ergic 53, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Syntaxin ergic 53
    Phosphorylation of Sec16 is linked to ERES number. (A–C) Hells cells were transfected with cDNA encoding wild-type GFP-Sec16 (GFP–Sec16-wt; A) or GFP–Sec16-T415I (B), fixed 24 h later, and immunostained for Sec31 to label ERESs or <t>ERGIC-53</t> to label the ERGIC. (C) The number of ERESs and peripheral ERGIC punctae was counted. The bar graph represents the number of punctae positive for Sec31 (ERES) or ERGIC-53 (ERGIC). The values determined in cells expressing GFP–Sec16-wt were set to 100% (means ± SD; three independent experiments; asterisks indicate statistically significant difference to GFP–Sec16-wt; *, P
    Ergic 53, supplied by Syntaxin, used in various techniques. Bioz Stars score: 89/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology lman1 ergic 53 antibody
    Phosphorylation of Sec16 is linked to ERES number. (A–C) Hells cells were transfected with cDNA encoding wild-type GFP-Sec16 (GFP–Sec16-wt; A) or GFP–Sec16-T415I (B), fixed 24 h later, and immunostained for Sec31 to label ERESs or <t>ERGIC-53</t> to label the ERGIC. (C) The number of ERESs and peripheral ERGIC punctae was counted. The bar graph represents the number of punctae positive for Sec31 (ERES) or ERGIC-53 (ERGIC). The values determined in cells expressing GFP–Sec16-wt were set to 100% (means ± SD; three independent experiments; asterisks indicate statistically significant difference to GFP–Sec16-wt; *, P
    Lman1 Ergic 53 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti ergic 53
    Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): <t>anti-ERGIC-53;</t> Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm
    Anti Ergic 53, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore ergic 53 p58
    Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): <t>anti-ERGIC-53;</t> Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm
    Ergic 53 P58, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam α ergic 53
    Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): <t>anti-ERGIC-53;</t> Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm
    α Ergic 53, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Axxora mouse anti ergic 53
    Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): <t>anti-ERGIC-53;</t> Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm
    Mouse Anti Ergic 53, supplied by Axxora, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology rabbit anti ergic 53
    Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): <t>anti-ERGIC-53;</t> Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm
    Rabbit Anti Ergic 53, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enzo Biochem mouse anti ergic 53
    Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): <t>anti-ERGIC-53;</t> Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm
    Mouse Anti Ergic 53, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 88/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen ergic 53 reporter constructs
    Binding of Sec24 isoforms to transport signals in vitro . <t>ERGIC-53</t> tail peptides carrying the indicated motifs were coupled to thiol-activated Sepharose and the beads were incubated with precleared HeLa cell lysate. Bound proteins were separated by SDS–polyacrylamide
    Ergic 53 Reporter Constructs, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enzo Biochem mouse ergic 53
    Binding of Sec24 isoforms to transport signals in vitro . <t>ERGIC-53</t> tail peptides carrying the indicated motifs were coupled to thiol-activated Sepharose and the beads were incubated with precleared HeLa cell lysate. Bound proteins were separated by SDS–polyacrylamide
    Mouse Ergic 53, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rabbit ergic 53
    Binding of Sec24 isoforms to transport signals in vitro . <t>ERGIC-53</t> tail peptides carrying the indicated motifs were coupled to thiol-activated Sepharose and the beads were incubated with precleared HeLa cell lysate. Bound proteins were separated by SDS–polyacrylamide
    Rabbit Ergic 53, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    PLN arginine mutants are mislocalized out of the ER. ( A ) HEK cells were transiently transfected with tagged WT-PLN or PLN mutants and visualized by confocal microscopy to detect the flag tag. Note that mutation of either Arg13 or Arg14, or both together, resulted in mislocalization of PLN out of the ER with significant expression at the plasma membrane. Cells shown represent at least 20 representative cells per condition. ( B ) Primary mouse fibroblasts were transiently transfected with tagged WT-PLN or the RΔ14 PLN construct and visualized by confocal microscopy. Endogenous ER markers ERGIC-53 and calreticulin as well as plasma membrane protein markers Na/K ATPase and Na/Ca exchanger (NCX) were probed against to allow accurate visualization of the ER and the plasma membrane. WT-PLN shows staining similar to the ER markers and RΔ14 PLN shows more diffuse ER staining as well as pronounced plasma membrane staining. Inserts shows a higher magnification of the cell membrane. Note, no plasma membrane staining is seen in WT-PLN and the other ER proteins but clear membrane staining is seen for RΔ14 PLN and the plasma membrane proteins NCX and Na/K ATPase. (Scale bar, 10 µm)

    Journal: PLoS ONE

    Article Title: Endoplasmic Reticulum Protein Targeting of Phospholamban: A Common Role for an N-Terminal Di-Arginine Motif in ER Retention?

    doi: 10.1371/journal.pone.0011496

    Figure Lengend Snippet: PLN arginine mutants are mislocalized out of the ER. ( A ) HEK cells were transiently transfected with tagged WT-PLN or PLN mutants and visualized by confocal microscopy to detect the flag tag. Note that mutation of either Arg13 or Arg14, or both together, resulted in mislocalization of PLN out of the ER with significant expression at the plasma membrane. Cells shown represent at least 20 representative cells per condition. ( B ) Primary mouse fibroblasts were transiently transfected with tagged WT-PLN or the RΔ14 PLN construct and visualized by confocal microscopy. Endogenous ER markers ERGIC-53 and calreticulin as well as plasma membrane protein markers Na/K ATPase and Na/Ca exchanger (NCX) were probed against to allow accurate visualization of the ER and the plasma membrane. WT-PLN shows staining similar to the ER markers and RΔ14 PLN shows more diffuse ER staining as well as pronounced plasma membrane staining. Inserts shows a higher magnification of the cell membrane. Note, no plasma membrane staining is seen in WT-PLN and the other ER proteins but clear membrane staining is seen for RΔ14 PLN and the plasma membrane proteins NCX and Na/K ATPase. (Scale bar, 10 µm)

    Article Snippet: Other antibodies were obtained commercially: FLAG antibody (M2)(Sigma), His antibody (Qiagen), COP I β subunit (Abcam), NCX (Santa Cruz), Na/K ATPase (Hybridoma Bank), SERCA2a (Affinity Bioreagents), ERGIC 53 (Sigma), Calnexin (Abcam), Calreticulin (Affinity Bioreagents), PDI (Calbiochem), Estrogen Receptor beta (Abcam), ARFGAP (Santa Cruz), ARF1 (Abcam), SOAT antibody (Abcam), TGN46 (AbD Serotec) a kind gift from Dr Walter Kahr (U of Toronto), Alexa mouse Fluor488 and rabbit Fluor633 (Invitrogen).

    Techniques: Transfection, Confocal Microscopy, FLAG-tag, Mutagenesis, Expressing, Construct, Staining

    TANGO1 amino acids 1255–1295 are the minimal TEER. ( A ) A schematic depiction of myc-epitope tagged mitochondrially-targeted (mit-TEER) truncates. ( B ) mit-TEER truncates were expressed in 2H5 cells, fixed and stained with anti-myc-antibody and visualised with confocal microscopy. ( C ) mit-TEER truncates were expressed in 2H5 cells, which were fixed and stained using anti-myc antibody (green) and, as a mitochondrial marker, anti-HSP60 antibody (red). ( D ) Overlap of the signal from myc and HSP60 was quantified and plotted as the Manders’ overlap coefficient for the two constructs (mit-Δ1255–1295 and mit-Δ1296–1336 respectively). ( E ) 2H5 cells were transfected with mit-Δ1255–1295 or mit-Δ1296–1336, fixed, and stained with anti-myc, anti-HSP60 and anti-ERGIC-53 antibodies. Arrows indicate myc staining with or without colocalised ERGIC-53 staining. ( F ) The extent of overlap of ERGIC-53 and myc was quantified and plotted as the Manders’ overlap coefficient for mit-Δ1255–1295 and mit-Δ1296–1336, respectively. Scale bars: ( B, C, E and F ) 20 μm; inset 2 μm.

    Journal: eLife

    Article Title: TANGO1 builds a machine for collagen export by recruiting and spatially organizing COPII, tethers and membranes

    doi: 10.7554/eLife.32723

    Figure Lengend Snippet: TANGO1 amino acids 1255–1295 are the minimal TEER. ( A ) A schematic depiction of myc-epitope tagged mitochondrially-targeted (mit-TEER) truncates. ( B ) mit-TEER truncates were expressed in 2H5 cells, fixed and stained with anti-myc-antibody and visualised with confocal microscopy. ( C ) mit-TEER truncates were expressed in 2H5 cells, which were fixed and stained using anti-myc antibody (green) and, as a mitochondrial marker, anti-HSP60 antibody (red). ( D ) Overlap of the signal from myc and HSP60 was quantified and plotted as the Manders’ overlap coefficient for the two constructs (mit-Δ1255–1295 and mit-Δ1296–1336 respectively). ( E ) 2H5 cells were transfected with mit-Δ1255–1295 or mit-Δ1296–1336, fixed, and stained with anti-myc, anti-HSP60 and anti-ERGIC-53 antibodies. Arrows indicate myc staining with or without colocalised ERGIC-53 staining. ( F ) The extent of overlap of ERGIC-53 and myc was quantified and plotted as the Manders’ overlap coefficient for mit-Δ1255–1295 and mit-Δ1296–1336, respectively. Scale bars: ( B, C, E and F ) 20 μm; inset 2 μm.

    Article Snippet: Reviewer #1: TANGO1 interacts with CTAGE5 and COPII components Sec23/Sec24 and recruits ERGIC-53 containing membranes to generate a mega-transport carrier for export of collagens from the ER.

    Techniques: Staining, Confocal Microscopy, Marker, Construct, Transfection

    The NRZ tether links TANGO1 to ERGIC membranes. ( A ) Rings of TANGO1 (green) in RDEB/FB/C7 cells with RINT1 (red) and ERGIC-53 (blue). Deconvolved z-stacks of ten cells were used to quantify the location of the tether protein RINT1 relative to a ring of TANGO1. 90 rings of TANGO1 were manually scored, three adjacent slices in the image stack were used to identify signal from RINT1 in the vicinity of the ring of TANGO1. 23 rings showed RINT1 within the ring, 19 rings showed RINT1 on the circumference (at the edge) of the ring, 39 had RINT1 outside the ring, nine rings showed no detectable RINT1. ( B ) TANGO1, TANGO1Δ1255–1295, TANGO1Δ1296–1336 and TANGO1-Lum were expressed in HEK293T cells and immunoprecipitated. Samples were probed for NBAS, RINT1, cTAGE5 and ZW10. TANGO1 and TANGO1Δ1296–1336 immunoprecipitated all four proteins, but TANGO1Δ1255–1295 did not immunoprecipitate tether proteins. TANGO1-Lum did not pull down any of the four proteins. ( C ) RDEB/FB/C7 were transfected with siRNA (siCTRL, siNBAS, siRINT1 and siTANGO1) and immunostained for intracellular collagen VII (red) and calreticulin (green). ( D ) Quantification of fluorescence associated with intracellular collagen VII in ( C ). ( E ) Collagen VII secreted by RDEB/FB/C7 was looked at as the ratio of collagen in the medium to the lysate, quantified, and plotted as the average of values from at least three independent experiments. β-actin is a loading control. ( F ) siRINT1-, siNBAS- and siTANGO1-treated RDEB/FB/C7 were stained for collagen VII and ERGIC-53. ( G ) A plot of Manders’ overlap coefficient for ERGIC-53 and collagen VII from ( F ) used to quantify ERGIC-53 localisation to collagen accumulations. ( H ) Representative blots showing the efficiency of knockdown of NBAS, RINT1 and ZW10, quantified and plotted as the average ±s.d. from at least three independent experiments. ***p

    Journal: eLife

    Article Title: TANGO1 builds a machine for collagen export by recruiting and spatially organizing COPII, tethers and membranes

    doi: 10.7554/eLife.32723

    Figure Lengend Snippet: The NRZ tether links TANGO1 to ERGIC membranes. ( A ) Rings of TANGO1 (green) in RDEB/FB/C7 cells with RINT1 (red) and ERGIC-53 (blue). Deconvolved z-stacks of ten cells were used to quantify the location of the tether protein RINT1 relative to a ring of TANGO1. 90 rings of TANGO1 were manually scored, three adjacent slices in the image stack were used to identify signal from RINT1 in the vicinity of the ring of TANGO1. 23 rings showed RINT1 within the ring, 19 rings showed RINT1 on the circumference (at the edge) of the ring, 39 had RINT1 outside the ring, nine rings showed no detectable RINT1. ( B ) TANGO1, TANGO1Δ1255–1295, TANGO1Δ1296–1336 and TANGO1-Lum were expressed in HEK293T cells and immunoprecipitated. Samples were probed for NBAS, RINT1, cTAGE5 and ZW10. TANGO1 and TANGO1Δ1296–1336 immunoprecipitated all four proteins, but TANGO1Δ1255–1295 did not immunoprecipitate tether proteins. TANGO1-Lum did not pull down any of the four proteins. ( C ) RDEB/FB/C7 were transfected with siRNA (siCTRL, siNBAS, siRINT1 and siTANGO1) and immunostained for intracellular collagen VII (red) and calreticulin (green). ( D ) Quantification of fluorescence associated with intracellular collagen VII in ( C ). ( E ) Collagen VII secreted by RDEB/FB/C7 was looked at as the ratio of collagen in the medium to the lysate, quantified, and plotted as the average of values from at least three independent experiments. β-actin is a loading control. ( F ) siRINT1-, siNBAS- and siTANGO1-treated RDEB/FB/C7 were stained for collagen VII and ERGIC-53. ( G ) A plot of Manders’ overlap coefficient for ERGIC-53 and collagen VII from ( F ) used to quantify ERGIC-53 localisation to collagen accumulations. ( H ) Representative blots showing the efficiency of knockdown of NBAS, RINT1 and ZW10, quantified and plotted as the average ±s.d. from at least three independent experiments. ***p

    Article Snippet: Reviewer #1: TANGO1 interacts with CTAGE5 and COPII components Sec23/Sec24 and recruits ERGIC-53 containing membranes to generate a mega-transport carrier for export of collagens from the ER.

    Techniques: Immunoprecipitation, Transfection, Fluorescence, Staining

    F805C selectively co-localizes with BAP31. (A) Representative confocal images of HEK293 cells expressing F805C-Kv11.1 protein showing the overlap between anti-Kv11.1 and anti-ER/ERGIC protein immunostaining. Anti-Kv11.1 immunostaining is shown in red; anti-calnexin ( n = 12 images), anti-Sec31 ( n = 19 images), anti-ERGIC-53 ( n = 19 images), anti-derlin-1 ( n = 18 images), anti-KDEL ( n = 15 images), or anti-BAP31 ( n = 17 images) is shown as green; overlapping immunostaining of red and green signals of similar intensities is shown as yellow; and the cell nuclei are labeled blue. The arrowheads highlight the distinct immunostaining pattern of F805C-Kv11.1 protein, and the scale bar represents 10 μm. (B) The mean Costes’ Pearson Coefficient (PC) for the image data sets are shown. PC values at or below the dashed line are (PC = 0.5) indicative of weak or no co-localization. Two independent cultures were tested for each condition.

    Journal: Frontiers in Physiology

    Article Title: Visualizing Mutation-Specific Differences in the Trafficking-Deficient Phenotype of Kv11.1 Proteins Linked to Long QT Syndrome Type 2

    doi: 10.3389/fphys.2018.00584

    Figure Lengend Snippet: F805C selectively co-localizes with BAP31. (A) Representative confocal images of HEK293 cells expressing F805C-Kv11.1 protein showing the overlap between anti-Kv11.1 and anti-ER/ERGIC protein immunostaining. Anti-Kv11.1 immunostaining is shown in red; anti-calnexin ( n = 12 images), anti-Sec31 ( n = 19 images), anti-ERGIC-53 ( n = 19 images), anti-derlin-1 ( n = 18 images), anti-KDEL ( n = 15 images), or anti-BAP31 ( n = 17 images) is shown as green; overlapping immunostaining of red and green signals of similar intensities is shown as yellow; and the cell nuclei are labeled blue. The arrowheads highlight the distinct immunostaining pattern of F805C-Kv11.1 protein, and the scale bar represents 10 μm. (B) The mean Costes’ Pearson Coefficient (PC) for the image data sets are shown. PC values at or below the dashed line are (PC = 0.5) indicative of weak or no co-localization. Two independent cultures were tested for each condition.

    Article Snippet: The cells were incubated with 2 mL blocking solution (10% goat serum in PBS) for 1 h to block non-specific binding sites, and the cells were subsequently incubated with the appropriate primary antibodies: anti-Kv11.1 (1:1000, Alomone Laboratories, Jerusalem, Israel), anti-calnexin (1:1000, Abcam, Cambridge, MA, United States), anti-KDEL (1:1000, Abcam, Cambridge, MA, United States), anti-Sec31a (1:100, BD Labs, Franklin Lakes, NJ, United States), anti-Bap31 (1:500, Abcam, Cambridge, MA, United States), and anti-ERGIC-53 (1:1000, Alexis Biochemicals, Plymouth Meeting, PA, United States) mixed with blocking solution at room temperature.

    Techniques: Expressing, Immunostaining, Labeling

    TFG accumulates at the ER/ERGIC interface in mammalian cells The distributions of endogenous TFG and ERGIC-53 were examined in hTERT-immortalized RPE-1 cells stably expressing low levels of mApple-Sec16B using SR-SIM (more than 15 cells analyzed on 3

    Journal: The EMBO Journal

    Article Title: TFG clusters COPII-coated transport carriers and promotes early secretory pathway organization

    doi: 10.15252/embj.201489032

    Figure Lengend Snippet: TFG accumulates at the ER/ERGIC interface in mammalian cells The distributions of endogenous TFG and ERGIC-53 were examined in hTERT-immortalized RPE-1 cells stably expressing low levels of mApple-Sec16B using SR-SIM (more than 15 cells analyzed on 3

    Article Snippet: Staining using antibodies directed against ERGIC-53 (Enzo Life Sciences, ALX-804-602), Sec31A (BD Biosciences, 612351), ERp57 (Proteintech, 15967-1-AP), GM130 (BD Biosciences, 610822) and TFG antibodies was typically preceded by fixation using 4% paraformaldehyde at 37°C, except in cases where cells were counterstained using Sec16A or β-COP antibodies.

    Techniques: Stable Transfection, Expressing

    Analysis of nocodazole induced Golgi fragmentation in mSar1p injected cells. Vero cells were incubated for 30 min at 4°C to depolymerize microtubules. Cells were microinjected in the cold with 2 mg/ml purified mSar1p mixed with 2 mg/ml cascade blue conjugated BSA. Immediately after injection, cells were incubated for 1 h ( a and b ) or 2 h ( c and d ) at 37°C in the presence of nocodazole (10 μM) before they were fixed and stained for ERGIC 53 ( a , c , and e ) or the Golgi marker giantin ( b , d , and f ). In e and f , cells were first treated with nocodazole for 2 h, microinjected and subsequently incubated in the presence of nocodazole for 1 h. Microinjected cells (marked by asterisks) were identified by the coinjected BSA (not shown). Microinjection of mSar1p results in the accumulation of ERGIC 53 in the ER but does not interfere with the formation and distribution of the Golgi fragments (marked by arrowheads in b ); in noninjected cells ERGIC 53 and giantin often appear to colocalize in peripheral Golgi fragments (marked by arrows). Bar, 15 μm.

    Journal: The Journal of Cell Biology

    Article Title: An Ordered Inheritance Strategy for the Golgi Apparatus: Visualization of Mitotic Disassembly Reveals a Role for the Mitotic Spindle

    doi:

    Figure Lengend Snippet: Analysis of nocodazole induced Golgi fragmentation in mSar1p injected cells. Vero cells were incubated for 30 min at 4°C to depolymerize microtubules. Cells were microinjected in the cold with 2 mg/ml purified mSar1p mixed with 2 mg/ml cascade blue conjugated BSA. Immediately after injection, cells were incubated for 1 h ( a and b ) or 2 h ( c and d ) at 37°C in the presence of nocodazole (10 μM) before they were fixed and stained for ERGIC 53 ( a , c , and e ) or the Golgi marker giantin ( b , d , and f ). In e and f , cells were first treated with nocodazole for 2 h, microinjected and subsequently incubated in the presence of nocodazole for 1 h. Microinjected cells (marked by asterisks) were identified by the coinjected BSA (not shown). Microinjection of mSar1p results in the accumulation of ERGIC 53 in the ER but does not interfere with the formation and distribution of the Golgi fragments (marked by arrowheads in b ); in noninjected cells ERGIC 53 and giantin often appear to colocalize in peripheral Golgi fragments (marked by arrows). Bar, 15 μm.

    Article Snippet: Antibodies for these experiments include: affinity-purified rabbit antisera to rat GM130 ( ); affinity-purified rabbit antisera to human giantin ( ); antisera recognizing the ER marker p62 ( ); monoclonal anti-ERGIC 53 ( ); monoclonal anti–α-tubulin (clone B512) from Sigma Chemical Co. ; monoclonal GTL2 raised against rat β-1,4 galactosyltransferase ( ).

    Techniques: Injection, Incubation, Purification, Staining, Marker

    Transport inhibition by a dominant mutant of Sar1p (mSar1p). ( a ) Schematic representation of the Golgi-to-ER recycling model for Golgi disassembly, and the experimental approach using mSar1p to test this model. ( b–e ) Vero cells were microinjected with 2 mg/ml purified mSar1p mixed with 2 mg/ml cascade blue-conjugated BSA which served as a coinjection marker (not shown). After incubation of cells for 20 min at 37°C, they were injected a second time with plasmids encoding the plasma membrane marker CD8 or NAGFP and incubated as described in experimental procedures. ( b ) NAGFP-specific fluorescence; ( c ) staining for CD8; ( d ) staining for endogenous ERGIC 53. In cells injected with mSar1p (asterisks) all markers analyzed accumulated in the ER. In noninjected cells CD8 and NAGFP were transported to the plasma membrane or Golgi complex (marked by an arrow in b ), respectively, and ERGIC 53 showed a typical distribution of the intermediate compartment. Arrowheads in b and c point to the nuclear envelope staining which is characteristic for ER localization. In e , mSar1p-injected (asterisk) and control cells were treated with 5 μg/ml BFA for 20 min, the BFA was washed out and cells fixed 2 h later and stained for the endogenous Golgi marker, giantin. In the presence of mSar1p, exit from the ER was inhibited for the Golgi resident giantin. Bars: ( b , c , and e ) 15 μm; ( d ) 10 μm.

    Journal: The Journal of Cell Biology

    Article Title: An Ordered Inheritance Strategy for the Golgi Apparatus: Visualization of Mitotic Disassembly Reveals a Role for the Mitotic Spindle

    doi:

    Figure Lengend Snippet: Transport inhibition by a dominant mutant of Sar1p (mSar1p). ( a ) Schematic representation of the Golgi-to-ER recycling model for Golgi disassembly, and the experimental approach using mSar1p to test this model. ( b–e ) Vero cells were microinjected with 2 mg/ml purified mSar1p mixed with 2 mg/ml cascade blue-conjugated BSA which served as a coinjection marker (not shown). After incubation of cells for 20 min at 37°C, they were injected a second time with plasmids encoding the plasma membrane marker CD8 or NAGFP and incubated as described in experimental procedures. ( b ) NAGFP-specific fluorescence; ( c ) staining for CD8; ( d ) staining for endogenous ERGIC 53. In cells injected with mSar1p (asterisks) all markers analyzed accumulated in the ER. In noninjected cells CD8 and NAGFP were transported to the plasma membrane or Golgi complex (marked by an arrow in b ), respectively, and ERGIC 53 showed a typical distribution of the intermediate compartment. Arrowheads in b and c point to the nuclear envelope staining which is characteristic for ER localization. In e , mSar1p-injected (asterisk) and control cells were treated with 5 μg/ml BFA for 20 min, the BFA was washed out and cells fixed 2 h later and stained for the endogenous Golgi marker, giantin. In the presence of mSar1p, exit from the ER was inhibited for the Golgi resident giantin. Bars: ( b , c , and e ) 15 μm; ( d ) 10 μm.

    Article Snippet: Antibodies for these experiments include: affinity-purified rabbit antisera to rat GM130 ( ); affinity-purified rabbit antisera to human giantin ( ); antisera recognizing the ER marker p62 ( ); monoclonal anti-ERGIC 53 ( ); monoclonal anti–α-tubulin (clone B512) from Sigma Chemical Co. ; monoclonal GTL2 raised against rat β-1,4 galactosyltransferase ( ).

    Techniques: Inhibition, Mutagenesis, Purification, Marker, Incubation, Injection, Fluorescence, Staining

    Phosphorylation of Sec16 is linked to ERES number. (A–C) Hells cells were transfected with cDNA encoding wild-type GFP-Sec16 (GFP–Sec16-wt; A) or GFP–Sec16-T415I (B), fixed 24 h later, and immunostained for Sec31 to label ERESs or ERGIC-53 to label the ERGIC. (C) The number of ERESs and peripheral ERGIC punctae was counted. The bar graph represents the number of punctae positive for Sec31 (ERES) or ERGIC-53 (ERGIC). The values determined in cells expressing GFP–Sec16-wt were set to 100% (means ± SD; three independent experiments; asterisks indicate statistically significant difference to GFP–Sec16-wt; *, P

    Journal: The Journal of Cell Biology

    Article Title: MAPK signaling to the early secretory pathway revealed by kinase/phosphatase functional screening

    doi: 10.1083/jcb.200912082

    Figure Lengend Snippet: Phosphorylation of Sec16 is linked to ERES number. (A–C) Hells cells were transfected with cDNA encoding wild-type GFP-Sec16 (GFP–Sec16-wt; A) or GFP–Sec16-T415I (B), fixed 24 h later, and immunostained for Sec31 to label ERESs or ERGIC-53 to label the ERGIC. (C) The number of ERESs and peripheral ERGIC punctae was counted. The bar graph represents the number of punctae positive for Sec31 (ERES) or ERGIC-53 (ERGIC). The values determined in cells expressing GFP–Sec16-wt were set to 100% (means ± SD; three independent experiments; asterisks indicate statistically significant difference to GFP–Sec16-wt; *, P

    Article Snippet: To show that the budding assay for ERGIC-53 is COPII dependent, we performed budding assays with cytosol immunodepleted for Sec24A and -B ( ).

    Techniques: Transfection, Expressing

    ERK2 stimulates COPII vesicle budding. HeLa microsomes were incubated for 30 min with ATP and an ATP-regenerating system, GTP, and cytosol of ERK2-silenced HeLa cells in the presence or absence of purified ERK2. COPII vesicles (V) were collected by centrifugation and immunoblotted for anti–ERGIC-53 and anti–CLIMP-63 (ER marker). The asterisk represents a statistically significant difference (*, P

    Journal: The Journal of Cell Biology

    Article Title: MAPK signaling to the early secretory pathway revealed by kinase/phosphatase functional screening

    doi: 10.1083/jcb.200912082

    Figure Lengend Snippet: ERK2 stimulates COPII vesicle budding. HeLa microsomes were incubated for 30 min with ATP and an ATP-regenerating system, GTP, and cytosol of ERK2-silenced HeLa cells in the presence or absence of purified ERK2. COPII vesicles (V) were collected by centrifugation and immunoblotted for anti–ERGIC-53 and anti–CLIMP-63 (ER marker). The asterisk represents a statistically significant difference (*, P

    Article Snippet: To show that the budding assay for ERGIC-53 is COPII dependent, we performed budding assays with cytosol immunodepleted for Sec24A and -B ( ).

    Techniques: Incubation, Purification, Centrifugation, Marker

    Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): anti-ERGIC-53; Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm

    Journal: Cellular and Molecular Life Sciences

    Article Title: Direct evidence that the N-terminal extensions of the TAP complex act as autonomous interaction scaffolds for the assembly of the MHC I peptide-loading complex

    doi: 10.1007/s00018-012-1005-6

    Figure Lengend Snippet: Subcellular localization of TMD 0 TAP1 and TMD 0 TAP2 in tapasin deficient cells. TMD 0 TAP1 ( a ), TMD 0 TAP2 ( b ), and wt TAP1/2 ( c ) were transiently expressed in the tapasin deficient cell line M553, stained with the antibodies as indicated in Fig. 2 , and analyzed via indirect immunofluorescence. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): anti-ERGIC-53; Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm

    Article Snippet: As organelle makers, rabbit anti-Calreticulin (polyclonal IgG fraction; Sigma-Aldrich, Steinheim, Germany), anti-ERGIC-53 (Sigma-Aldrich), and anti-GM130 (Golgi ; EP892Y; Abcam) were used.

    Techniques: Staining, Immunofluorescence, Marker

    Subcellular localization of TMD 0 TAP1/2 . Indirect immunofluorescence on transiently transfected HeLa cells (TMD 0 s or wt TAP1/2 as a control) was performed using TMD 0 TAP1 , TMD 0 TAP2 and TAP1 specific (myc, HA and 148.3, respectively, green ) as well as organelle specific antibodies [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): ERGIC-53; Golgi GM-130; red ]. a TMD 0 TAP1 , b TMD 0 TAP2 , c wt TAP. Images were taken using confocal laser scanning microscopy. Pearson coefficients were calculated from 8–10 individual images. Scale bar 10 μm

    Journal: Cellular and Molecular Life Sciences

    Article Title: Direct evidence that the N-terminal extensions of the TAP complex act as autonomous interaction scaffolds for the assembly of the MHC I peptide-loading complex

    doi: 10.1007/s00018-012-1005-6

    Figure Lengend Snippet: Subcellular localization of TMD 0 TAP1/2 . Indirect immunofluorescence on transiently transfected HeLa cells (TMD 0 s or wt TAP1/2 as a control) was performed using TMD 0 TAP1 , TMD 0 TAP2 and TAP1 specific (myc, HA and 148.3, respectively, green ) as well as organelle specific antibodies [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): ERGIC-53; Golgi GM-130; red ]. a TMD 0 TAP1 , b TMD 0 TAP2 , c wt TAP. Images were taken using confocal laser scanning microscopy. Pearson coefficients were calculated from 8–10 individual images. Scale bar 10 μm

    Article Snippet: As organelle makers, rabbit anti-Calreticulin (polyclonal IgG fraction; Sigma-Aldrich, Steinheim, Germany), anti-ERGIC-53 (Sigma-Aldrich), and anti-GM130 (Golgi ; EP892Y; Abcam) were used.

    Techniques: Immunofluorescence, Transfection, Confocal Laser Scanning Microscopy

    Subcellular localization of coreTAP. HeLa cells were transiently transfected with coreTAP1-eGFP ( a ), coreTAP2-mCerulean ( b ), or a combination of both coreTAP constructs ( c ), and analyzed by confocal laser scanning microscopy. CoreTAP2-mCerulean was indirectly visualized via its C-terminal StrepII-tag with a StrepII antibody. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): anti-ERGIC-53; Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm

    Journal: Cellular and Molecular Life Sciences

    Article Title: Direct evidence that the N-terminal extensions of the TAP complex act as autonomous interaction scaffolds for the assembly of the MHC I peptide-loading complex

    doi: 10.1007/s00018-012-1005-6

    Figure Lengend Snippet: Subcellular localization of coreTAP. HeLa cells were transiently transfected with coreTAP1-eGFP ( a ), coreTAP2-mCerulean ( b ), or a combination of both coreTAP constructs ( c ), and analyzed by confocal laser scanning microscopy. CoreTAP2-mCerulean was indirectly visualized via its C-terminal StrepII-tag with a StrepII antibody. Organelles were stained with antibodies against marker proteins [ ER anti-calreticulin; ER- Golg i intermediate compartment ( ERGIC ): anti-ERGIC-53; Golgi : anti-GM130]. Pearson coefficients were calculated from 8 to 10 individual images. Scale bar 10 μm

    Article Snippet: As organelle makers, rabbit anti-Calreticulin (polyclonal IgG fraction; Sigma-Aldrich, Steinheim, Germany), anti-ERGIC-53 (Sigma-Aldrich), and anti-GM130 (Golgi ; EP892Y; Abcam) were used.

    Techniques: Transfection, Construct, Confocal Laser Scanning Microscopy, Staining, Marker

    Binding of Sec24 isoforms to transport signals in vitro . ERGIC-53 tail peptides carrying the indicated motifs were coupled to thiol-activated Sepharose and the beads were incubated with precleared HeLa cell lysate. Bound proteins were separated by SDS–polyacrylamide

    Journal: EMBO Reports

    Article Title: Role of Sec24 isoforms in selective export of membrane proteins from the endoplasmic reticulum

    doi: 10.1038/sj.embor.7400893

    Figure Lengend Snippet: Binding of Sec24 isoforms to transport signals in vitro . ERGIC-53 tail peptides carrying the indicated motifs were coupled to thiol-activated Sepharose and the beads were incubated with precleared HeLa cell lysate. Bound proteins were separated by SDS–polyacrylamide

    Article Snippet: After 24 h, the cells were transfected with ERGIC-53 reporter constructs (Effectene, Qiagen).

    Techniques: Binding Assay, In Vitro, Incubation

    Sec24D overexpression does not compensate for the loss of Sec24A and Sec24B. ( A , B ) HeLa cells were transfected with siRNA against Sec24A ( A ) or Sec24A and Sec24B ( B ). After 1 day, the cells were co-transfected with Myc–Sec24D and Myc–ERGIC-53

    Journal: EMBO Reports

    Article Title: Role of Sec24 isoforms in selective export of membrane proteins from the endoplasmic reticulum

    doi: 10.1038/sj.embor.7400893

    Figure Lengend Snippet: Sec24D overexpression does not compensate for the loss of Sec24A and Sec24B. ( A , B ) HeLa cells were transfected with siRNA against Sec24A ( A ) or Sec24A and Sec24B ( B ). After 1 day, the cells were co-transfected with Myc–Sec24D and Myc–ERGIC-53

    Article Snippet: After 24 h, the cells were transfected with ERGIC-53 reporter constructs (Effectene, Qiagen).

    Techniques: Over Expression, Transfection

    Effects of Sec24 isoform-specific knockdown on signal-mediated transport. ( A ) A Myc-tagged glycosylated variant of human ERGIC-53 was used as a reporter for endoplasmic reticulum to Golgi transport. The −1 and −2 positions (XX) in the

    Journal: EMBO Reports

    Article Title: Role of Sec24 isoforms in selective export of membrane proteins from the endoplasmic reticulum

    doi: 10.1038/sj.embor.7400893

    Figure Lengend Snippet: Effects of Sec24 isoform-specific knockdown on signal-mediated transport. ( A ) A Myc-tagged glycosylated variant of human ERGIC-53 was used as a reporter for endoplasmic reticulum to Golgi transport. The −1 and −2 positions (XX) in the

    Article Snippet: After 24 h, the cells were transfected with ERGIC-53 reporter constructs (Effectene, Qiagen).

    Techniques: Variant Assay