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Combination of CXB/DMC/SD‐133 with CDH11 activates the cyclic GMP‐AMP synthase‐stimulator of the interferon gene (cGAS‐STING) pathway through ROS, suppressing cellular invasion and migration. A) Representative western blots of the cellular thermal shift assay showing increase in CDH11 thermostable performance in the presence of CXB, DMC, and SD‐133. B) SD‐133 with interactive residue side chains at the pocket are shown in stick rendering, with the inhibitors drawn in colorful. The polypeptide backbones are rendered as ribbons. The yellow broken lines indicate potential intermolecular hydrogen bonds, while the gray broken lines indicate pi‐cation interactions. C–E) Effect of CXB, DMC, and SD‐133 on the inhibition of SACC‐83 cells; normalized data and non‐linear regression curve fitting are shown. IC 50 values are indicated. F) BCAA per million SACC‐83 cells exhibited a decrease 24 h after treatment with CXB, DMC, or SD‐133. Mean ± SEM is shown, * P < 0.05 using t test. G) SACC‐83 cells treated with CXB, DMC, or SD‐133 analyzed for their migration and invasion ability using transwell assays. Scale bar, 100 µm, n = 3, Mean ± SEM is shown, * P < 0.05 using t test. H) Top 10 terms of GO enrichment analysis of DEG by RNA‐seq in SACC‐83 cells treated with SD133. I) Expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated with CXB, DMC, and SD 133, assessed using western blotting. J) Flow cytometry reveals an increase in reactive oxygen species (ROS) production after treatment with CXB, DMC, or SD‐133. K,L) Confocal microscopy showing the accumulation and quantification of cytosolic deoxyribonucleic acid (DNA) in SACC‐83 cells following treatment with CXB, DMC, or SD‐133. Double‐stranded DNA (dsDNA) visualized using <t>PicoGreen</t> <t>staining</t> (green), while MitoTracker (red) and <t>DAPI</t> (blue) employed to label mitochondria and nuclei, respectively. Scale bar, 5 µm. More than 100 cells were analyzed per group. n = 10, Means ± SEM is shown. *** P < 0.001 using one‐way analysis of variance. M) Representative images of DNA comet assays of SACC‐83 cells subjected to treatment with CXB, DMC, or SD‐133.
Dapi Staining Solution 9556, supplied by ZSGB Biotech, 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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Combination of CXB/DMC/SD‐133 with CDH11 activates the cyclic GMP‐AMP synthase‐stimulator of the interferon gene (cGAS‐STING) pathway through ROS, suppressing cellular invasion and migration. A) Representative western blots of the cellular thermal shift assay showing increase in CDH11 thermostable performance in the presence of CXB, DMC, and SD‐133. B) SD‐133 with interactive residue side chains at the pocket are shown in stick rendering, with the inhibitors drawn in colorful. The polypeptide backbones are rendered as ribbons. The yellow broken lines indicate potential intermolecular hydrogen bonds, while the gray broken lines indicate pi‐cation interactions. C–E) Effect of CXB, DMC, and SD‐133 on the inhibition of SACC‐83 cells; normalized data and non‐linear regression curve fitting are shown. IC 50 values are indicated. F) BCAA per million SACC‐83 cells exhibited a decrease 24 h after treatment with CXB, DMC, or SD‐133. Mean ± SEM is shown, * P < 0.05 using t test. G) SACC‐83 cells treated with CXB, DMC, or SD‐133 analyzed for their migration and invasion ability using transwell assays. Scale bar, 100 µm, n = 3, Mean ± SEM is shown, * P < 0.05 using t test. H) Top 10 terms of GO enrichment analysis of DEG by RNA‐seq in SACC‐83 cells treated with SD133. I) Expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated with CXB, DMC, and SD 133, assessed using western blotting. J) Flow cytometry reveals an increase in reactive oxygen species (ROS) production after treatment with CXB, DMC, or SD‐133. K,L) Confocal microscopy showing the accumulation and quantification of cytosolic deoxyribonucleic acid (DNA) in SACC‐83 cells following treatment with CXB, DMC, or SD‐133. Double‐stranded DNA (dsDNA) visualized using PicoGreen staining (green), while MitoTracker (red) and DAPI (blue) employed to label mitochondria and nuclei, respectively. Scale bar, 5 µm. More than 100 cells were analyzed per group. n = 10, Means ± SEM is shown. *** P < 0.001 using one‐way analysis of variance. M) Representative images of DNA comet assays of SACC‐83 cells subjected to treatment with CXB, DMC, or SD‐133.

Journal: Advanced Science

Article Title: Inhibition of CDH11 Activates cGAS‐STING by Stimulating Branched Chain Amino Acid Catabolism and Mitigates Lung Metastasis of Adenoid Cystic Carcinoma

doi: 10.1002/advs.202408751

Figure Lengend Snippet: Combination of CXB/DMC/SD‐133 with CDH11 activates the cyclic GMP‐AMP synthase‐stimulator of the interferon gene (cGAS‐STING) pathway through ROS, suppressing cellular invasion and migration. A) Representative western blots of the cellular thermal shift assay showing increase in CDH11 thermostable performance in the presence of CXB, DMC, and SD‐133. B) SD‐133 with interactive residue side chains at the pocket are shown in stick rendering, with the inhibitors drawn in colorful. The polypeptide backbones are rendered as ribbons. The yellow broken lines indicate potential intermolecular hydrogen bonds, while the gray broken lines indicate pi‐cation interactions. C–E) Effect of CXB, DMC, and SD‐133 on the inhibition of SACC‐83 cells; normalized data and non‐linear regression curve fitting are shown. IC 50 values are indicated. F) BCAA per million SACC‐83 cells exhibited a decrease 24 h after treatment with CXB, DMC, or SD‐133. Mean ± SEM is shown, * P < 0.05 using t test. G) SACC‐83 cells treated with CXB, DMC, or SD‐133 analyzed for their migration and invasion ability using transwell assays. Scale bar, 100 µm, n = 3, Mean ± SEM is shown, * P < 0.05 using t test. H) Top 10 terms of GO enrichment analysis of DEG by RNA‐seq in SACC‐83 cells treated with SD133. I) Expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated with CXB, DMC, and SD 133, assessed using western blotting. J) Flow cytometry reveals an increase in reactive oxygen species (ROS) production after treatment with CXB, DMC, or SD‐133. K,L) Confocal microscopy showing the accumulation and quantification of cytosolic deoxyribonucleic acid (DNA) in SACC‐83 cells following treatment with CXB, DMC, or SD‐133. Double‐stranded DNA (dsDNA) visualized using PicoGreen staining (green), while MitoTracker (red) and DAPI (blue) employed to label mitochondria and nuclei, respectively. Scale bar, 5 µm. More than 100 cells were analyzed per group. n = 10, Means ± SEM is shown. *** P < 0.001 using one‐way analysis of variance. M) Representative images of DNA comet assays of SACC‐83 cells subjected to treatment with CXB, DMC, or SD‐133.

Article Snippet: Subsequently, the cells were rinsed thrice with PBS and stained with DAPI Staining Solution (ZSGB‐BIO, 9556, China).

Techniques: Migration, Western Blot, Thermal Shift Assay, Residue, Inhibition, RNA Sequencing, Expressing, Flow Cytometry, Confocal Microscopy, Staining

Metabolism of BCAA increases the production of ROS, activating the cGAS‐STING pathway. A) Flow cytometry detected ROS production in SACC‐83 cells with CDH11 knockdown. B) Representative images of DNA comet assays of SACC‐83 cells subjected to various experimental conditions. Scale bar, 20 µm. C,D) Confocal microscopy showing the accumulation and quantification of cytosolic DNA in SACC‐83 cells under knockdown CDH11. dsDNA visualized using PicoGreen staining (green), while MitoTracker (red) and DAPI (blue) employed to label mitochondria and nuclei, respectively. Scale bar, 5 µm. More than 100 cells were analyzed per group. Mean ± SEM is shown. n = 10, *** P < 0.001 using t test. E) Expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated under various experimental conditions, assessed using western blotting. F) Flow cytometry of SACC‐83 cells overexpressing CDH11 to evaluate the mitochondrial activity. G) Representative images of DNA comet assays of SACC‐83 cells subjected to various experimental conditions. Scale bar, 20 µm. H) Expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated under various experimental conditions, assessed using western blotting. I) qRT‐PCR analysis shows that CDH11‐knockdown cells significantly increase the expression of IFNB1. No significant difference in the overexpression group. n = 3, Mean ± SEM is shown, * P < 0.05 using t test. J) Flow cytometry detected ROS production in BCAA‐ deprived or ‐added SACC‐83 cells. K,L) Confocal microscopy showing the accumulation and quantification of cytosolic DNA in BCAA‐deprived or ‐added SACC‐83 cells. dsDNA visualized using PicoGreen staining (green), while MitoTracker (red) and DAPI (blue) employed to label mitochondria and nuclei, respectively. Scale bar, 5 µm. More than 100 cells were analyzed per group. n = 10, Mean ± SEM is shown. *** P < 0.001 using one‐way analysis of variance. M) The expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated under various experimental conditions, was assessed using western blotting.

Journal: Advanced Science

Article Title: Inhibition of CDH11 Activates cGAS‐STING by Stimulating Branched Chain Amino Acid Catabolism and Mitigates Lung Metastasis of Adenoid Cystic Carcinoma

doi: 10.1002/advs.202408751

Figure Lengend Snippet: Metabolism of BCAA increases the production of ROS, activating the cGAS‐STING pathway. A) Flow cytometry detected ROS production in SACC‐83 cells with CDH11 knockdown. B) Representative images of DNA comet assays of SACC‐83 cells subjected to various experimental conditions. Scale bar, 20 µm. C,D) Confocal microscopy showing the accumulation and quantification of cytosolic DNA in SACC‐83 cells under knockdown CDH11. dsDNA visualized using PicoGreen staining (green), while MitoTracker (red) and DAPI (blue) employed to label mitochondria and nuclei, respectively. Scale bar, 5 µm. More than 100 cells were analyzed per group. Mean ± SEM is shown. n = 10, *** P < 0.001 using t test. E) Expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated under various experimental conditions, assessed using western blotting. F) Flow cytometry of SACC‐83 cells overexpressing CDH11 to evaluate the mitochondrial activity. G) Representative images of DNA comet assays of SACC‐83 cells subjected to various experimental conditions. Scale bar, 20 µm. H) Expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated under various experimental conditions, assessed using western blotting. I) qRT‐PCR analysis shows that CDH11‐knockdown cells significantly increase the expression of IFNB1. No significant difference in the overexpression group. n = 3, Mean ± SEM is shown, * P < 0.05 using t test. J) Flow cytometry detected ROS production in BCAA‐ deprived or ‐added SACC‐83 cells. K,L) Confocal microscopy showing the accumulation and quantification of cytosolic DNA in BCAA‐deprived or ‐added SACC‐83 cells. dsDNA visualized using PicoGreen staining (green), while MitoTracker (red) and DAPI (blue) employed to label mitochondria and nuclei, respectively. Scale bar, 5 µm. More than 100 cells were analyzed per group. n = 10, Mean ± SEM is shown. *** P < 0.001 using one‐way analysis of variance. M) The expression of cGAS‐STING pathway‐related proteins in SACC‐83 cells, treated under various experimental conditions, was assessed using western blotting.

Article Snippet: Subsequently, the cells were rinsed thrice with PBS and stained with DAPI Staining Solution (ZSGB‐BIO, 9556, China).

Techniques: Flow Cytometry, Knockdown, Confocal Microscopy, Staining, Expressing, Western Blot, Activity Assay, Quantitative RT-PCR, Over Expression