Fig. 2. CeHop participates in the development of the gonads. (a) Coomassie-stained gel (lane 1 + lane 2) and Western blot analysis against recombinant His6-CeHop and endogenous Hop (lane 3 + lane 4). Recombinant protein was loaded in lanes 1 and 3, whereas whole worm extracts were loaded in lanes 2 and 4. The antibody was raised against CeHop and used as outlined in Materials and Methods. (b) The ectopic expression of EYFP under the control of the CeHop promoter is predominant in intestinal tissues and can be detected in several other tissues at lower levels. Tissues are labeled as Pm (pharyngeal muscle cells), Bwm (body-wall muscle cells), Sp (spermathecae), and Int (intestinal cell ring). (c) Light microscopy image of adult
hsf-1(
sy441)/R09E12.3(RNAi) and hsf
(sy441)/control(RNAi) worms. (d-g) DAPI staining of the gonads was performed in N2 and
hsf-1(
sy441) nematodes treated either with control RNAi or with RNAi against CeHop (R09E12.3). While
hsf-1(
sy441)/control (d) and N2/R09E12.3(RNAi) (g) developed normally,
hsf-1(
sy441)/R09E12.3(RNAi) developed malformed gonad arms (e; arrows) or no gonad arms (f; arrows). (h) DAPI staining of
hsf-1(
sy441)/R09E12.3(RNAi) germline cells. Oocytes emerging from the distal gonad show bivalent chromosomes (arrows). Spermatheca is marked with a triangle.