JCS02811 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Western blot analysis shows that b-tubulin is present in a Mr ~140,000 band that co-purifies with radial spokes and spoke stalks. The top panels show western blots of sucrose density gradient fractions of axonemal extracts from wild-type (WT) and the mutant pf17 probed with an antibody to RSP16, which serves as a marker for 20S radial spokes from wild-type cells and 15S spoke stalks from the mutant. The lower panels show western blots of the same fractions probed with an anti-sea-urchin b-tubulin antibody (Sigma T0198). The antibody recognized a 140 kDa band that co-sedimented with the 20S radial spokes from wild-type and 15S spoke stalks from pf17. As expected, the antibody also recognized a 55 kDa band corresponding to monomeric b-tubulin. Note that the 140 kDa band is discrete, and not the result of upward smearing of monomeric tubulin due to gel overloading.