Figure 5.

(A) Western blot of total cell lysate of wild-type cells (lane 1) and GFP-DdLim cells (lane 2) probed with affinity-purified anti-DdLim antibodies shows the monospecificity of these antibodies used for immunolocalization. GFP-DdLim fusion protein (lane 2) is expressed in these cells to an amount equal to that of the endogenous level of DdLim. (B) Immunolocalization of DdLim in wild-type cells using affinty-purified anti-DdLim antibodies. The phase contrast images are indicated by a, b, and c, the corresponding fluorescence images by a′, b′, and c′. Vegetative (a′ and b′), and preaggregation-competent cells (c′), were fixed with cold methanol and stained with 10 μg/ml of the anti-DdLim antibodies as the first antibody and TRITC-conjugated antirabbit as the second antibody. Scale bar, 10 μm.