Fig. 3. Immunoblots to measure expression of profilin-I-EGFP in Acanthamoeba. Proteins were detected with antibodies to profilin, actin or GFP by chemiluminescence. T, amoeba transfected with pUbPG and selected from pools M2 and M3; C, control untransfected amoeba. Actin at 43 kDa served as the loading control for these samples. Endogenous profilins run at 13 kDa. The 42 kDa band that reacts with both anti-profilin and anti-GFP is the fusion protein. The band at 44 kDa that reacts with anti-GFP in both lanes T and C is a crossreactive amoeba protein. The minor 27 kDa band that reacted with anti-GFP but not anti-profilin is likely to be proteolytic fragment of EGFP.