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mutant allele of cdk4 (i12a)  (Broad Institute Inc)

 
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    Broad Institute Inc mutant allele of cdk4 (i12a)
    (A) HFFs were split into media with palbociclib and BrdU incorporation was measured in cells harvested after 24 hours (3 bio rep). Student’s T test was calculated compared to 0 nM. (B) 200 nM palbociclib was added at times after split and BrdU incorporation was measured in cells harvested at 24 hours post split (2 bio rep). Student’s T-test was calculated compared to 0 hr. (C) HFFs were split into palbociclib and harvested at indicated times and analyzed by immunoblot. * indicates nonspecific band. (D) Parental HFFs were transduced with <t>pLenti-E1Fa-CDK4</t> WT, K35M, and <t>I12A</t> and contact arrested before harvested for immunoblot. Arrow indicates on-target band. (E) CDK4 expressing HFFs were released from contact arrest into palbociclib and harvested after 24 hours and analyzed by immunoblot. (F) CDK4 expressing HFFs were released from contact arrest into palbociclib and transcript level of CDC6 and MCM5 was measured by RT-qPCR (3 bio rep). Student’s T-test was calculated by comparing to 0 nM. (G) CDK4 expressing HFFs were released from contact arrest into palbociclib and S phase was measured by BrdU incorporation after 24 hours. Statistics comparing curves was completed by two-way ANOVA (3 bio rep). P values are indicated as * for <0.05, and ** for <0.01, *** for <0.001. See also .
    Mutant Allele Of Cdk4 (I12a), supplied by Broad Institute Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mutant+allele+of+cdk4+(i12a)/crispr+guide+sequences+against+cdk4/pmc06586519-81-22-37
    Average 90 stars, based on 1 article reviews
    mutant allele of cdk4 (i12a) - by Bioz Stars, 2026-11
    90/100 stars

    Images

    1) Product Images from "Cyclin D-CDK4 relieves cooperative repression of proliferation and cell cycle gene expression by DREAM and RB."

    Article Title: Cyclin D-CDK4 relieves cooperative repression of proliferation and cell cycle gene expression by DREAM and RB.

    Journal: Oncogene

    doi: 10.1038/s41388-019-0767-9

    (A) HFFs were split into media with palbociclib and BrdU incorporation was measured in cells harvested after 24 hours (3 bio rep). Student’s T test was calculated compared to 0 nM. (B) 200 nM palbociclib was added at times after split and BrdU incorporation was measured in cells harvested at 24 hours post split (2 bio rep). Student’s T-test was calculated compared to 0 hr. (C) HFFs were split into palbociclib and harvested at indicated times and analyzed by immunoblot. * indicates nonspecific band. (D) Parental HFFs were transduced with pLenti-E1Fa-CDK4 WT, K35M, and I12A and contact arrested before harvested for immunoblot. Arrow indicates on-target band. (E) CDK4 expressing HFFs were released from contact arrest into palbociclib and harvested after 24 hours and analyzed by immunoblot. (F) CDK4 expressing HFFs were released from contact arrest into palbociclib and transcript level of CDC6 and MCM5 was measured by RT-qPCR (3 bio rep). Student’s T-test was calculated by comparing to 0 nM. (G) CDK4 expressing HFFs were released from contact arrest into palbociclib and S phase was measured by BrdU incorporation after 24 hours. Statistics comparing curves was completed by two-way ANOVA (3 bio rep). P values are indicated as * for <0.05, and ** for <0.01, *** for <0.001. See also .
    Figure Legend Snippet: (A) HFFs were split into media with palbociclib and BrdU incorporation was measured in cells harvested after 24 hours (3 bio rep). Student’s T test was calculated compared to 0 nM. (B) 200 nM palbociclib was added at times after split and BrdU incorporation was measured in cells harvested at 24 hours post split (2 bio rep). Student’s T-test was calculated compared to 0 hr. (C) HFFs were split into palbociclib and harvested at indicated times and analyzed by immunoblot. * indicates nonspecific band. (D) Parental HFFs were transduced with pLenti-E1Fa-CDK4 WT, K35M, and I12A and contact arrested before harvested for immunoblot. Arrow indicates on-target band. (E) CDK4 expressing HFFs were released from contact arrest into palbociclib and harvested after 24 hours and analyzed by immunoblot. (F) CDK4 expressing HFFs were released from contact arrest into palbociclib and transcript level of CDC6 and MCM5 was measured by RT-qPCR (3 bio rep). Student’s T-test was calculated by comparing to 0 nM. (G) CDK4 expressing HFFs were released from contact arrest into palbociclib and S phase was measured by BrdU incorporation after 24 hours. Statistics comparing curves was completed by two-way ANOVA (3 bio rep). P values are indicated as * for <0.05, and ** for <0.01, *** for <0.001. See also .

    Techniques Used: BrdU Incorporation Assay, Western Blot, Transduction, Expressing, Quantitative RT-PCR



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    mutant allele of cdk4 (i12a)  (Broad Institute Inc)
    90
    Broad Institute Inc mutant allele of cdk4 (i12a)
    (A) HFFs were split into media with palbociclib and BrdU incorporation was measured in cells harvested after 24 hours (3 bio rep). Student’s T test was calculated compared to 0 nM. (B) 200 nM palbociclib was added at times after split and BrdU incorporation was measured in cells harvested at 24 hours post split (2 bio rep). Student’s T-test was calculated compared to 0 hr. (C) HFFs were split into palbociclib and harvested at indicated times and analyzed by immunoblot. * indicates nonspecific band. (D) Parental HFFs were transduced with <t>pLenti-E1Fa-CDK4</t> WT, K35M, and <t>I12A</t> and contact arrested before harvested for immunoblot. Arrow indicates on-target band. (E) CDK4 expressing HFFs were released from contact arrest into palbociclib and harvested after 24 hours and analyzed by immunoblot. (F) CDK4 expressing HFFs were released from contact arrest into palbociclib and transcript level of CDC6 and MCM5 was measured by RT-qPCR (3 bio rep). Student’s T-test was calculated by comparing to 0 nM. (G) CDK4 expressing HFFs were released from contact arrest into palbociclib and S phase was measured by BrdU incorporation after 24 hours. Statistics comparing curves was completed by two-way ANOVA (3 bio rep). P values are indicated as * for <0.05, and ** for <0.01, *** for <0.001. See also .
    Mutant Allele Of Cdk4 (I12a), supplied by Broad Institute Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mutant+allele+of+cdk4+(i12a)/crispr+guide+sequences+against+cdk4/pmc06586519-81-22-37
    Average 90 stars, based on 1 article reviews
    mutant allele of cdk4 (i12a) - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) HFFs were split into media with palbociclib and BrdU incorporation was measured in cells harvested after 24 hours (3 bio rep). Student’s T test was calculated compared to 0 nM. (B) 200 nM palbociclib was added at times after split and BrdU incorporation was measured in cells harvested at 24 hours post split (2 bio rep). Student’s T-test was calculated compared to 0 hr. (C) HFFs were split into palbociclib and harvested at indicated times and analyzed by immunoblot. * indicates nonspecific band. (D) Parental HFFs were transduced with pLenti-E1Fa-CDK4 WT, K35M, and I12A and contact arrested before harvested for immunoblot. Arrow indicates on-target band. (E) CDK4 expressing HFFs were released from contact arrest into palbociclib and harvested after 24 hours and analyzed by immunoblot. (F) CDK4 expressing HFFs were released from contact arrest into palbociclib and transcript level of CDC6 and MCM5 was measured by RT-qPCR (3 bio rep). Student’s T-test was calculated by comparing to 0 nM. (G) CDK4 expressing HFFs were released from contact arrest into palbociclib and S phase was measured by BrdU incorporation after 24 hours. Statistics comparing curves was completed by two-way ANOVA (3 bio rep). P values are indicated as * for <0.05, and ** for <0.01, *** for <0.001. See also .

    Journal: Oncogene

    Article Title: Cyclin D-CDK4 relieves cooperative repression of proliferation and cell cycle gene expression by DREAM and RB.

    doi: 10.1038/s41388-019-0767-9

    Figure Lengend Snippet: (A) HFFs were split into media with palbociclib and BrdU incorporation was measured in cells harvested after 24 hours (3 bio rep). Student’s T test was calculated compared to 0 nM. (B) 200 nM palbociclib was added at times after split and BrdU incorporation was measured in cells harvested at 24 hours post split (2 bio rep). Student’s T-test was calculated compared to 0 hr. (C) HFFs were split into palbociclib and harvested at indicated times and analyzed by immunoblot. * indicates nonspecific band. (D) Parental HFFs were transduced with pLenti-E1Fa-CDK4 WT, K35M, and I12A and contact arrested before harvested for immunoblot. Arrow indicates on-target band. (E) CDK4 expressing HFFs were released from contact arrest into palbociclib and harvested after 24 hours and analyzed by immunoblot. (F) CDK4 expressing HFFs were released from contact arrest into palbociclib and transcript level of CDC6 and MCM5 was measured by RT-qPCR (3 bio rep). Student’s T-test was calculated by comparing to 0 nM. (G) CDK4 expressing HFFs were released from contact arrest into palbociclib and S phase was measured by BrdU incorporation after 24 hours. Statistics comparing curves was completed by two-way ANOVA (3 bio rep). P values are indicated as * for <0.05, and ** for <0.01, *** for <0.001. See also .

    Article Snippet: To determine if palbociclib-mediated inhibition of cell cycle progression was dependent on CDK4 activity in HFFs, we utilized a mutant allele of CDK4 (I12A) that is partially resistant to palbociclib (personal communication, Nicole Persky and Cory Johannessen, Broad Institute).

    Techniques: BrdU Incorporation Assay, Western Blot, Transduction, Expressing, Quantitative RT-PCR