d609 (cs-0078) Search Results


d609 (cs-0078)  (CEM Corporation)
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NT-EqtII exhibits a specific affinity for sphingomyelin located in the cytosolic leaflet of the living iMEF cell PM. ( a ) Schematic representation of NT-Eqt II observation in the cytosolic leaflet of the cell PM. ( b ) Fluorescent images of NT-EqtII-HaloTag7 labeled with SF650B, mEos4b-STING, and mEos4b-Rab6a in iMEFs. The images were acquired by oblique illumination (upper) and TIRF microscopy (TIRFM) (lower). Single fluorescent spots of NT-EqtII are indicated by yellow arrowheads. Scale bars, 2 μm. ( c ) Fluorescent images of NT-EqtII-HaloTag7 labeled with SF650B in various cellular contexts, encompassing wild-type (WT) iMEFs, SMS1 and 2-DKO iMEFs, iMEFs expressing WT bacterial SMase or its catalytically inactive mutant harboring Ras farnesylation sequence by which is anchored to the cytosolic leaflet of the PM, along with iMEFs subjected to <t>D609</t> treatment. The images were acquired by oblique illumination (left) and TIRFM (right). Quantification of NT-EqtII-HaloTag7 expressed in cells was performed by evaluating fluorescence intensity via oblique illumination. The content of the SM probe in the cytosolic leaflet of the cell PM was quantified by measuring the numbers of single spots (yellow arrowheads) by TIRFM. Scale bars, 2 μm. ( d ) The numbers of localizations of NT-EqtII-HaloTag7 labeled with SF650B bound to the cytosolic leaflets of cell PMs, were determined by TIRFM at 4 ms/frame for 2000 frames and subsequently normalized by the expressed amounts (fluorescence intensities in the cytoplasm minus those in the background) measured by oblique illumination. The value in the vertical axis shows the difference between the numbers of localizations of NT-EqtII-Halo7-SF650B in cells expressing NT-EqtII and those in cells that do not express NT-EqtII but were incubated with SF650B. The presented error bars symbolize standard errors. *** and n.s. indicate significant (p < 0.001) and not significant differences (p > 0.05), respectively, using Welch’s T -test.
D609 (Cs 0078), supplied by CEM Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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NT-EqtII exhibits a specific affinity for sphingomyelin located in the cytosolic leaflet of the living iMEF cell PM. ( a ) Schematic representation of NT-Eqt II observation in the cytosolic leaflet of the cell PM. ( b ) Fluorescent images of NT-EqtII-HaloTag7 labeled with SF650B, mEos4b-STING, and mEos4b-Rab6a in iMEFs. The images were acquired by oblique illumination (upper) and TIRF microscopy (TIRFM) (lower). Single fluorescent spots of NT-EqtII are indicated by yellow arrowheads. Scale bars, 2 μm. ( c ) Fluorescent images of NT-EqtII-HaloTag7 labeled with SF650B in various cellular contexts, encompassing wild-type (WT) iMEFs, SMS1 and 2-DKO iMEFs, iMEFs expressing WT bacterial SMase or its catalytically inactive mutant harboring Ras farnesylation sequence by which is anchored to the cytosolic leaflet of the PM, along with iMEFs subjected to D609 treatment. The images were acquired by oblique illumination (left) and TIRFM (right). Quantification of NT-EqtII-HaloTag7 expressed in cells was performed by evaluating fluorescence intensity via oblique illumination. The content of the SM probe in the cytosolic leaflet of the cell PM was quantified by measuring the numbers of single spots (yellow arrowheads) by TIRFM. Scale bars, 2 μm. ( d ) The numbers of localizations of NT-EqtII-HaloTag7 labeled with SF650B bound to the cytosolic leaflets of cell PMs, were determined by TIRFM at 4 ms/frame for 2000 frames and subsequently normalized by the expressed amounts (fluorescence intensities in the cytoplasm minus those in the background) measured by oblique illumination. The value in the vertical axis shows the difference between the numbers of localizations of NT-EqtII-Halo7-SF650B in cells expressing NT-EqtII and those in cells that do not express NT-EqtII but were incubated with SF650B. The presented error bars symbolize standard errors. *** and n.s. indicate significant (p < 0.001) and not significant differences (p > 0.05), respectively, using Welch’s T -test.

Journal: Scientific Reports

Article Title: A non-toxic equinatoxin-II reveals the dynamics and distribution of sphingomyelin in the cytosolic leaflet of the plasma membrane

doi: 10.1038/s41598-024-67803-2

Figure Lengend Snippet: NT-EqtII exhibits a specific affinity for sphingomyelin located in the cytosolic leaflet of the living iMEF cell PM. ( a ) Schematic representation of NT-Eqt II observation in the cytosolic leaflet of the cell PM. ( b ) Fluorescent images of NT-EqtII-HaloTag7 labeled with SF650B, mEos4b-STING, and mEos4b-Rab6a in iMEFs. The images were acquired by oblique illumination (upper) and TIRF microscopy (TIRFM) (lower). Single fluorescent spots of NT-EqtII are indicated by yellow arrowheads. Scale bars, 2 μm. ( c ) Fluorescent images of NT-EqtII-HaloTag7 labeled with SF650B in various cellular contexts, encompassing wild-type (WT) iMEFs, SMS1 and 2-DKO iMEFs, iMEFs expressing WT bacterial SMase or its catalytically inactive mutant harboring Ras farnesylation sequence by which is anchored to the cytosolic leaflet of the PM, along with iMEFs subjected to D609 treatment. The images were acquired by oblique illumination (left) and TIRFM (right). Quantification of NT-EqtII-HaloTag7 expressed in cells was performed by evaluating fluorescence intensity via oblique illumination. The content of the SM probe in the cytosolic leaflet of the cell PM was quantified by measuring the numbers of single spots (yellow arrowheads) by TIRFM. Scale bars, 2 μm. ( d ) The numbers of localizations of NT-EqtII-HaloTag7 labeled with SF650B bound to the cytosolic leaflets of cell PMs, were determined by TIRFM at 4 ms/frame for 2000 frames and subsequently normalized by the expressed amounts (fluorescence intensities in the cytoplasm minus those in the background) measured by oblique illumination. The value in the vertical axis shows the difference between the numbers of localizations of NT-EqtII-Halo7-SF650B in cells expressing NT-EqtII and those in cells that do not express NT-EqtII but were incubated with SF650B. The presented error bars symbolize standard errors. *** and n.s. indicate significant (p < 0.001) and not significant differences (p > 0.05), respectively, using Welch’s T -test.

Article Snippet: D609 (CS-0078) was purchased from CEM.

Techniques: Labeling, Microscopy, Expressing, Mutagenesis, Sequencing, Fluorescence, Incubation