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skco1  (ATCC)


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    ATCC skco1
    Validation of mutant selectivity of KRAS G12V siRNA in vitro (A) UV melting curves and sequences of 23mer duplexes between the fully modified KRAS guide RNA and the targeted G12V mutant, WT, and G12D mutant RNA. (B) Western blot analysis in A431 cells stably expressing KRAS WT or G12V transiently transfected with siRNAs. Cells were analyzed at 48 hours and 72 hours. Blots were done separately, and densitometry quantification below is based on vinculin control for each individual blot. (C) RT-qPCR analysis in A431 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 48 hours. Data shown as mean +/- SEM, experiments performed in duplicate. (D) Luciferase dose-response curve in A431-KRAS-WT or A431-KRAS-G12V cells stably expressing a luciferase reporter. Cells were analyzed at 72 hours. Data shown as mean +/- SEM, experiments performed in triplicate. (E) Volcano plots from RNA-sequencing in <t>SKCO1</t> cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 24 hours. This figure includes data published in Stanland et al and has received permission to be shown in this figure.
    Skco1, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Protocols to evaluate mutant specificity of an oncogene-targeting siRNA using orthogonal in vitro and in vivo approaches"

    Article Title: Protocols to evaluate mutant specificity of an oncogene-targeting siRNA using orthogonal in vitro and in vivo approaches

    Journal: STAR Protocols

    doi: 10.1016/j.xpro.2025.104323

    Validation of mutant selectivity of KRAS G12V siRNA in vitro (A) UV melting curves and sequences of 23mer duplexes between the fully modified KRAS guide RNA and the targeted G12V mutant, WT, and G12D mutant RNA. (B) Western blot analysis in A431 cells stably expressing KRAS WT or G12V transiently transfected with siRNAs. Cells were analyzed at 48 hours and 72 hours. Blots were done separately, and densitometry quantification below is based on vinculin control for each individual blot. (C) RT-qPCR analysis in A431 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 48 hours. Data shown as mean +/- SEM, experiments performed in duplicate. (D) Luciferase dose-response curve in A431-KRAS-WT or A431-KRAS-G12V cells stably expressing a luciferase reporter. Cells were analyzed at 72 hours. Data shown as mean +/- SEM, experiments performed in triplicate. (E) Volcano plots from RNA-sequencing in SKCO1 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 24 hours. This figure includes data published in Stanland et al and has received permission to be shown in this figure.
    Figure Legend Snippet: Validation of mutant selectivity of KRAS G12V siRNA in vitro (A) UV melting curves and sequences of 23mer duplexes between the fully modified KRAS guide RNA and the targeted G12V mutant, WT, and G12D mutant RNA. (B) Western blot analysis in A431 cells stably expressing KRAS WT or G12V transiently transfected with siRNAs. Cells were analyzed at 48 hours and 72 hours. Blots were done separately, and densitometry quantification below is based on vinculin control for each individual blot. (C) RT-qPCR analysis in A431 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 48 hours. Data shown as mean +/- SEM, experiments performed in duplicate. (D) Luciferase dose-response curve in A431-KRAS-WT or A431-KRAS-G12V cells stably expressing a luciferase reporter. Cells were analyzed at 72 hours. Data shown as mean +/- SEM, experiments performed in triplicate. (E) Volcano plots from RNA-sequencing in SKCO1 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 24 hours. This figure includes data published in Stanland et al and has received permission to be shown in this figure.

    Techniques Used: Biomarker Discovery, Mutagenesis, In Vitro, Modification, Western Blot, Stable Transfection, Expressing, Transfection, Control, Quantitative RT-PCR, Luciferase, RNA Sequencing



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    Validation of mutant selectivity of KRAS G12V siRNA in vitro (A) UV melting curves and sequences of 23mer duplexes between the fully modified KRAS guide RNA and the targeted G12V mutant, WT, and G12D mutant RNA. (B) Western blot analysis in A431 cells stably expressing KRAS WT or G12V transiently transfected with siRNAs. Cells were analyzed at 48 hours and 72 hours. Blots were done separately, and densitometry quantification below is based on vinculin control for each individual blot. (C) RT-qPCR analysis in A431 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 48 hours. Data shown as mean +/- SEM, experiments performed in duplicate. (D) Luciferase dose-response curve in A431-KRAS-WT or A431-KRAS-G12V cells stably expressing a luciferase reporter. Cells were analyzed at 72 hours. Data shown as mean +/- SEM, experiments performed in triplicate. (E) Volcano plots from RNA-sequencing in <t>SKCO1</t> cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 24 hours. This figure includes data published in Stanland et al and has received permission to be shown in this figure.
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    Image Search Results


    Validation of mutant selectivity of KRAS G12V siRNA in vitro (A) UV melting curves and sequences of 23mer duplexes between the fully modified KRAS guide RNA and the targeted G12V mutant, WT, and G12D mutant RNA. (B) Western blot analysis in A431 cells stably expressing KRAS WT or G12V transiently transfected with siRNAs. Cells were analyzed at 48 hours and 72 hours. Blots were done separately, and densitometry quantification below is based on vinculin control for each individual blot. (C) RT-qPCR analysis in A431 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 48 hours. Data shown as mean +/- SEM, experiments performed in duplicate. (D) Luciferase dose-response curve in A431-KRAS-WT or A431-KRAS-G12V cells stably expressing a luciferase reporter. Cells were analyzed at 72 hours. Data shown as mean +/- SEM, experiments performed in triplicate. (E) Volcano plots from RNA-sequencing in SKCO1 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 24 hours. This figure includes data published in Stanland et al and has received permission to be shown in this figure.

    Journal: STAR Protocols

    Article Title: Protocols to evaluate mutant specificity of an oncogene-targeting siRNA using orthogonal in vitro and in vivo approaches

    doi: 10.1016/j.xpro.2025.104323

    Figure Lengend Snippet: Validation of mutant selectivity of KRAS G12V siRNA in vitro (A) UV melting curves and sequences of 23mer duplexes between the fully modified KRAS guide RNA and the targeted G12V mutant, WT, and G12D mutant RNA. (B) Western blot analysis in A431 cells stably expressing KRAS WT or G12V transiently transfected with siRNAs. Cells were analyzed at 48 hours and 72 hours. Blots were done separately, and densitometry quantification below is based on vinculin control for each individual blot. (C) RT-qPCR analysis in A431 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 48 hours. Data shown as mean +/- SEM, experiments performed in duplicate. (D) Luciferase dose-response curve in A431-KRAS-WT or A431-KRAS-G12V cells stably expressing a luciferase reporter. Cells were analyzed at 72 hours. Data shown as mean +/- SEM, experiments performed in triplicate. (E) Volcano plots from RNA-sequencing in SKCO1 cells transiently transfected with siRNAs at 20 nM. Cells were analyzed at 24 hours. This figure includes data published in Stanland et al and has received permission to be shown in this figure.

    Article Snippet: SKCO1 , ATCC , Cat #HTB-39; RRID: CVCL_0626.

    Techniques: Biomarker Discovery, Mutagenesis, In Vitro, Modification, Western Blot, Stable Transfection, Expressing, Transfection, Control, Quantitative RT-PCR, Luciferase, RNA Sequencing