Figure 3.

Effects of culture conditions on PDE4D3 localization and migration. (A) L6-C5 cells were cultured for 24 h in 1% BSA or 1% FBS. After harvesting the cells, soluble, Triton-X-100–soluble and –insoluble fractions were prepared as described in MATERIALS AND METHODS and submitted to SDS-PAGE. Approximately 50 μg of protein were loaded in each lane. PDE4D3 was detected by using the M3S1 antibody. A representative experiment of the four performed is reported. (B) L6-C5 myoblasts were transiently transfected with pCMV5-PDE4D3 plasmid, cultured, and treated as described in MATERIALS AND METHODS. Immunofluorescence was performed after 24 h of culture in DMEM containing 1% BSA or 1% FBS as indicated, using M3S1 as first antibody.