Journal: FEBS letters

Article Title: Overexpression of Mos(rat) proto-oncogene product enhances the positive autoregulatory loop of MyoD.

doi: 10.1016/s0014-5793(98)01192-2

Figure Lengend Snippet: Fig. 4. Mos co-expressed with MyoD activates expression of the endogenous MyoD protein. 10T1/2 cells were co-transfected with 0.5 Wg of pEMSV-E12 (lanes 1^14), 0.2, 0.5 and 1 Wg of pCMVHA-MosWT (lanes 2^4 and 9^11) or pCMVHA-MosKM (lanes 5^7 and 12^14) alone or in combination with 0.5 Wg of pCMVHa-MyoD (lanes 8^14). Forty-eight hours following transfection, cells maintained in DMEM supplemented with 15% FCS were harvested and Western blot analysis was performed. A: Ten micrograms of whole cell extracts was solubilized in SDS loading bu¡er. After SDS-PAGE, HA-tagged proteins were detected by immunoblotting with the 12CA5 monoclonal antibody. B: Fifty micro- grams of the same whole extracts was used for SDS-PAGE. Proteins were detected by immunoblotting with the a¤nity-puri¢ed polyclonal anti-MyoD antibody.

Article Snippet: After electrophoretic transfer of proteins onto nitrocellulose membranes, immunodetection was performed with the 12CA5 monoclonal antibody (dilution: 1/1000, Boehringer) or the MyoD antibody (dilution: 1/1000, Santa Cruz).

Techniques: Expressing, Transfection, Western Blot, SDS Page