Figure 3 | Oncogene

Figure 3

From: Selective repression of myoD transcription by v-Myc prevents terminal differentiation of quail embryo myoblasts transformed by the MC29 strain of avian myelocytomatosis virus

Figure 3

Rescue of myogenic differentiation by exogenous MyoD in QM(myc). QM(myc) cells were super-infected with an empty (Ad-null) or a MyoD-carrying adenoviral vector under the control of Rous sarcoma virus LTR (Ad-MyoD) (Murry et al., 1996). Multiplicity of infection ranged between 100 and 1000. Differentiation Medium (F14 medium containing 2% FCS and 0.5 μg/ml bovine insulin) (DM) was added the day after infection and cells were analysed after further 3 days. (a) Western blot analysis using the anti-Myc polyclonal antibody 237 (Crouch et al., 1993), to reveal the P110gag-v-myc protein, the anti-skeletal muscle Myosin Heavy Chain (MHC) monoclonal antibody MF20, and an anti-Myogenin polyclonal antibody. (b) Ad-MyoD super-infected cells were fixed with 3% paraformaldehyde, permeabilized with 0.25% Triton X-100, and incubated with the polyclonal antiserum raised against murine MyoD expressed in bacteria R4B4 (Russo et al., 1997) and with the above referred antibodies against MHC and Myogenin. Nuclei were stained with the Hoechst 33258 dye (Sigma), that revealed also the abundant occurrence of apoptotic figures (A,D), as expected for myc-transformed cells in a low serum medium. Left micrographs represent the same field where MHC (B) and MyoD (C) were visualized. Right micrographs represent the same field, from a sister plate, where MHC (E) and Myogenin (F) expression were analysed. Bar=20 μm

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