Resistance to apoptosis conferred by Cdk inhibitors during myocyte differentiation

Science

Volumn 273, Issue 5273, 1996, Pages 359-361

  Wang, Jian ^a   Walsh, Kenneth ^a  

^a TUFTS UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

APOPTOSIS; ARTICLE; CELL DIFFERENTIATION; CELL PROLIFERATION; ENZYME INHIBITION; MITOSIS; MUSCLE CELL; MYOTUBE; PRIORITY JOURNAL; TISSUE SPECIFICITY;

EID: 0029996726     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5273.359     Document Type: Article

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28. C2C12 myocytes were maintained in Dulbecco's modified eagle's medium (DMEM) supplemented with fetal bovine Serum (20%, growth medium), and myogenic differentiation was initiated by shifting subconfluent cultures into differentiation medium [DMEM supplemented with horse serum (2%)]. For immunofluorescence microscopy, cells grown on sterile glass cover slips were fixed for 10 min with 2% paraformaldehyde (in phosphate-buffered saline) and permeabilized with 0.5% NP-40. After they were blocked for 5 min with bovine serum albumin (2%), cells were stained with a monoclonal antibody to skeletal MHC proteins (MF20) and rhodamine-conjugated antibody to mouse immunoglobulin G. The cells were then incubated with digoxigenin-dUTP terminal dioxynucleotide transferase mixture and subsequently stained with fluorescein-conjugated antibody to digoxigenin (ApopTag, Oncor), counter-stained with Hoechst 33258, and mounted. Specimens were examined and photographed with a Nikon Diaphot fluorescence microscope.
29. CIP1 recognizes both full-length p21 and p21 proteins truncated at the COOH-terminus. Monoclonal antibody to myogenin antibody was a gift of W. Wright [W. E. Wright, M. Binder, W. Funk, Mol. Cell. Biol. 11, 4104 (1991)].
30. INK4A, L. Zhu for pCMV-CD20, and W. Wright for antibody to myogenin. Supported by NIH grants HL50692 and AR40197.