Figure 6.

The ptl-1 deletion mutation does not affect overall microtubule structures in neurons. Adult wild type (N2) and ptl-1 null mutant worms were fixed and prepared for immunocytochemistry as described in Materials and methods. Freeze-cracked, fixed worms were incubated in mouse monoclonal antibody directed against tubulin (clone DM1α, Sigma) at 1:500, which is a general tubulin antibody (see text), and, following multiple washes, exposed to Alexa-fluor 488 nm goat anti-mouse secondary antibody (Molecular Probes, Inc) at 1:1000. Tubulin immunoreactivity was visualized using a PCM2000 Nikon confocal laser scanning microscope with an argon laser. A, C, F are wild type worms stained with the DM1 α antibody; B, D, E are ptl-1 mutant worms stained with the DM1 α antibody. G) acetylated tubulin (MEC-12; antibody 6-11-B1, 1:10) in ALML/R in ptl-1 mutant worm, H) ALML/R labeled with anti-acetylated tubulin (MEC-12) antibody in wild type neurons. The NR branch of ALM is visible in H and G. I) PLM neuron from ptl-1 mutant worm, labeled with anti-MEC-12 antibody. J) Control ptl-1 mutant worm with no primary antibody. Anterior is up in A, F, G, H, I and J. Anterior is to the right in B and E. Posterior is up in C and D.