Journal:

Article Title: Biallelic Mutations in p16 INK4a Confer Resistance to Ras- and Ets-Induced Senescence in Human Diploid Fibroblasts

doi: 10.1128/MCB.22.23.8135-8143.2002

Figure Lengend Snippet: Functional analyses of p16INK4a variants in Q34 cells. (A) Lysates from Q34 cells and the control fibroblasts, 904, previously infected with a retrovirus encoding simian virus 40 T antigen, were immunoprecipitated with polyclonal antibodies against Cdk4 or p16INK4a as indicated. Following SDS-12% PAGE the amounts of Cdk4 and p16INK4a in each precipitate were compared by immunoblotting with the corresponding monoclonal antibodies. IP, immunoprecipitation. (B) Longer exposures are shown of Cdk4 and Cdk6 immunoprecipitates that have been immunoblotted for p16INK4a and the respective Cdks. (C) Normal HDFs were infected with recombinant retroviruses encoding 2× HA-tagged p16INK4a (wild type [WT]), and the indicated variants and pools of infected cells were selected in puromycin. Seven days after infection, cell lysates were prepared and equivalent amounts (500 μg) of protein were immunoprecipitated with antibodies against Cdk4 or Cdk6 (upper two panels). Immunoprecipitated proteins were fractionated by SDS-12% PAGE and were immunoblotted with a monoclonal antibody against p16INK4a. The positions of exogenous (2×HA p16) and endogenous p16INK4a (End p16) are indicated. Samples (20 μg) of total lysate from the same cells were analyzed directly by SDS-PAGE and immunoblotting for p16INK4a and Cdk4 (lower panels). (D) Aliquots of each cell pool, at 7 days postselection, were transferred into 24-well plates (5 × 103 cells/well), and their proliferation was monitored for 10 days. At each time point cells were fixed in 10% formaldehyde and viable cells were stained with crystal violet. Relative numbers of cells were determined by measurements of the optical density at 590 nm. The data present the averages of triplicate measurements.

Article Snippet: The monoclonal antibody DO-1 against p53 (sc-126), the rabbit polyclonal antibodies against Cdk4 (sc-601) and Cdk6 (sc-177), and the SMP14 monoclonal against MDM2 (sc-965) were obtained from Santa Cruz.

Techniques: Functional Assay, Infection, Immunoprecipitation, Western Blot, Recombinant, SDS Page, Staining

Journal: Molecular and cellular endocrinology

Article Title: DUAL INHIBITION OF ERK1/2 AND AKT PATHWAYS IS REQUIRED TO SUPPRESS THE GROWTH AND SURVIVAL OF ENDOMETRIOTIC CELLS AND LESIONS

doi: 10.1016/j.mce.2018.12.011

Figure Lengend Snippet: Details of the antibody used.

Article Snippet: Anti-human mouse monoclonal CDK6 , Cell Signaling , 3136 , 1:1000.

Techniques: Concentration Assay