Proteins encoded by v-myc and c-myc oncogenes: Identification and localization in acute leukemia virus transformants and bursal lymphoma cell lines

Bin1 functionally interacts with Myc and inhibits cell proliferation via multiple mechanisms

“…Association of Myc and Bin1 was observed to coimmunoprecipitation from baculovirus-infected Sf9 cells and untransfected C2C12 myoblasts, where Bin1 function has been examined (Wechsler-Reya et al, 1998), using NP40 bu er conditions previously shown to support interaction of Myc and Bin1 in vitro (150 mM NaCl and 0.1% NP40). Bin1 was extracted more readily than Myc by NP40 lysis bu er from Sf9 cells infected with recombinant c-Myc and Bin1 baculoviruses, consistent with the fact that e cient extraction of Myc requires harsher conditions (RIPA bu er and sonication (Hann et al, 1983). However, the Myc complexes extracted under these conditions contained Bin1 as indicated by coimmunoprecipitation with Myc antibody (Figure 1a).…”

B-Myc is preferentially expressed in hormonally-controlled tissues and inhibits cellular proliferation

“…The fusion protein, GST-B-Myc50C, was expressed in BL21 E. Coli and puri®ed on glutathione agarose (Sigma) as described by Smith and Johnson (1988). Puri®ed GST-B-Myc50C was used for the generation of a nity-puri®ed rabbit polyclonal antiserum, anti-B-Myc50C, as previously described (Hann et al, 1983). The rabbit polyclonal antiserum, anti-MycN100 (against the N-terminal 100 amino acids of murine c-Myc), was generated as described previously (Lutterbach and Hann, 1994).…”

“…Cells were lysed in cold Ab lysis bu er (Hann et al, 1983) with 20 U/ml aprotinin (Sigma) followed by sonication. For radiolabeled cells, the amount of radiolabel incorporated into cellular proteins was determined by precipitation with 10% trichloroacetic acid (TCA).…”

“…Lysates were then incubated with 5 mg of the indicated antibody for 1 ± 2 h at 48C, and the immune complexes were precipitated with Pansorbin for 20 min. The immunoprecipitates were washed three times with cold RIPA bu er (Hann et al, 1983) and incubated at 958C for 3 min in Laemmli sample bu er. Samples were subjected to sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS ± PAGE) using 12.5 or 15% polyacrylamide gels followed by autoradiography.…”

“…Tissues dissected from adult ICR mice were homogenized and lysed in cold RIPA bu er (Hann et al, 1983) with 3% aprotinin and 0.1 mg/ml PMSF. Cultured cells were lysed in cold RIPA bu er.…”