Figure 2.

HuR and TRN2 are expressed during myogenesis, and associate until day 2 of differentiation. (A) Fifty, 100, and 300 ng of recombinant GST-TRN1 and GST-TRN2 were run on SDS-PAGE and subjected to Western blot analyses using antibodies against TRN1 and TRN2. SDS-PAGE followed by Coomassie staining of 1, 3, and 10 μg of recombinant protein demonstrates there was protein loaded in all lanes. (B) Western blot analyses were performed using 30 μg of total protein extracts from exponentially growing (Exp) and differentiating (day 0, d0, to day 4, d4) C2C12 cells. The blots were probed with antibodies to TRN1, TRN2, HuR, myoglobin, and β-actin. (C) Western blot analyses were performed using 100 μg of total protein extracts from exponentially growing (Exp) and differentiating (day 0, d0, to day 4, d4) C2C12 cells in order to detect the TRN1 protein. The blot was probed with antibodies to TRN1, myoglobin, and β-actin. (D) Total cell extracts were prepared from C2C12 cells on the indicated days during myogenesis and subjected to immunoprecipitation using a polyclonal α-TRN2 antibody. The immunoprecipitates were run on an SDS-PAGE, and a monoclonal anti-HuR antibody was used in Western blotting to look for association between the two proteins. HC refers to the heavy chain of the TRN2 antibody used for the IP.