Figure 7.

Disorganization of actin filaments in adult body wall muscle by RNA interference of Ce-kettin. (A) Protein levels of Ce-kettin in control or ketn-1 (RNAi) worms (50 worms per lane) in wild-type or rrf-3 background were examined by Western blot with MH44. The same membrane was reprobed with anti-myoA antibody to confirm equal loading of the proteins. (B) Frequency of formation of actin aggregates. Worms were stained with tetramethylrhodamine-phalloidin, and percentages of adult worms (n = 100) with actin aggregates in the body wall muscle were determined. (C–H) Actin organization in the body wall muscle of control RNAi (C, E, G, I, and K) or ketn-1 (RNAi) (D, F, H, J, and L) worms in wild-type (C, D, and I–L), rrf-3 (E and F), or unc-54 (G and H) backgrounds was examined by staining with tetramethylrhodamine-phalloidin. Worms in I–L were incubated in M9 buffer for 1 h in the absence (I and J) or presence (K and L) of 0.01% tetramisole before fixation. In ketn-1 (RNAi) worms, small round aggregates and unusual actin accumulations at the edges of muscle cells (arrows in D, F, and J) were detected, and this phenotype was enhanced by tetramisole-induced hypercontraction (L). Bar, 50 μm.