Journal: Molecular and Cellular Biology

Article Title: c-Myc Protein Synthesis Is Initiated from the Internal Ribosome Entry Segment during Apoptosis

doi: 10.1128/mcb.20.4.1162-1169.2000

Figure Lengend Snippet: FIG. 3. c-myc mRNA levels are unchanged during apoptosis. Poly(A)1 mRNA or total cellular RNA was prepared from samples taken over an 8-h period after the addition of TRAIL and then analyzed by Northern blotting for levels of c-myc mRNA, and levels of the control message GAPDH were determined using specific probes (A). The Northern blot was analyzed using a Molecular Dynamics PhosphorImager, and c-myc mRNA levels were normalized to those of GAPDH (B).

Article Snippet: Samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the amount of radiolabel incorporated was visualized on a PhosphorImager (Molecular Dynamics).

Techniques: Northern Blot, Control

Journal: Molecular and Cellular Biology

Article Title: c-Myc Protein Synthesis Is Initiated from the Internal Ribosome Entry Segment during Apoptosis

doi: 10.1128/mcb.20.4.1162-1169.2000

Figure Lengend Snippet: FIG. 4. c-Myc protein stability is unaltered during apoptosis. Determination of the half-life of the c-Myc protein in control HeLa cells and at 4 h post-addition of TRAIL is shown. Pulse-chase analysis and immunoprecipitation were per- formed as described in Materials and Methods. Cells were labeled for 30 min and then harvested following the chase times indicated. Samples were subjected to PAGE, and the amount of radiolabel incorporated into each band was deter- mined by using a phosphorimager. (A) Representative gels of pulse-chase– immunoprecipitations. (B) Phosphorimager analysis of the gels shown in panel A. Open circles, untreated HeLa cells; closed circles, HeLa cells incubated with TRAIL for 4 h.

Article Snippet: Samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the amount of radiolabel incorporated was visualized on a PhosphorImager (Molecular Dynamics).

Techniques: Control, Pulse Chase, Immunoprecipitation, Labeling, Incubation

Journal: Molecular and Cellular Biology

Article Title: Platelet-derived growth factor stimulation of GTPase-activating protein tyrosine phosphorylation in control and c-H-ras-expressing NIH 3T3 cells correlates with p21ras activation.

doi: 10.1128/mcb.12.9.3903

Figure Lengend Snippet: FIG. 2. Induction by PDGF of the tyrosine phosphorylation of a large percentage of cellular GAP. (A) Anti-GAP immunoblot analy- sis of PDGF-stimulated whole cell lysates. The amount of protein used per lane is noted at the bottom. (B) Anti-GAP immunoblot of anti-P-Tyr-recovered GAP from PDGF-stimulated cells. Aliquots of cell lysates containing the indicated amounts of protein were immu- noprecipitated with an excess amount of anti-P-Tyr (lanes la, 2a, and 3a). Following the first immunoprecipitation, fresh anti-P-Tyr was added to supernatants and the immunoprecipitation was re- peated (lanes lb, 2b, and 3b). It should be noted that >90% of the proteins recognized by anti-P-Tyr were recovered in the first immu- noprecipitation step. Comparison of the intensity of GAP in the anti-P-Tyr-recovered samples (a plus b lanes) with that in whole cell lysate blot was used to estimate the percentage of cellular GAP which was tyrosine phosphorylated. Anti-P-Tyr-recoverable GAP was calculated to be between 37 and 52% of total cellular GAP following quantitation of bound radioactivity by using a Molecular Dynamics model 400E Phosphorimager.

Article Snippet: Immunoreactive GAP was visualized by 12I-protein A and quantitated by using a Molecular Dynamics model 400E Phosphorimager.

Techniques: Phospho-proteomics, Western Blot, Immunoprecipitation, Comparison, Quantitation Assay, Radioactivity