spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.


Figure 6


Fig. 6. CrLis1 is absent from flagella lacking ODAs. (A) Wild-type flagella were electrophoresed and stained with Coomassie Blue (left panel) or blotted and probed with affinity-purified antibody raised against CrLis1 (right panel). The lower band at ~37 kDa comigrates with recombinant CrLis1 whereas the upper band (marked with an asterisk) is a cross-reacting protein that is completely missing in a mutant that lacks the central pair microtubule complex. (B) Flagella were demembranated with 1% IGEPAL-630 and the resulting axonemes extracted with 0.6 M NaCl. Equivalent amounts of each sample were electrophoresed in a 5-15% acrylamide gradient gel and stained with Coomassie Blue (upper panel) or blotted and probed with antibodies against CrLis1 and LC2 of the ODA (lower panels). Most CrLis1 is associated with axonemes and is extracted by high salt similar to LC2. However, we have observed that under different detergent conditions, more CrLis1 is present in the membrane plus matrix fraction. (C) Flagella from wild-type Chlamydomonas and mutants lacking various axonemal substructures were electrophoresed and stained with Coomassie Blue (upper panel) or immunoblotted to detect CrLis1 (lower panel). CrLis1 is specifically missing in strains lacking the ODAs (oda1 and oda2) or only the outer arm {alpha} HC and LC5 thiredoxin (oda11 and oda4-s7 oda11).





Right arrow Return to article