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. 2011 Oct 15;138(20):4387–4398. doi: 10.1242/dev.068098

Fig. 4.

Fig. 4.

Eya and Sall are required for protonephridia regeneration and Osr is expressed in tubule cells. (A) Eya(RNAi) animals failed to regenerate protonephridia. Cartoon depicts region analyzed. Left: numbers of tubule branches (Ac tubulin+) and tubule-associated cells (CA+) per cluster in control and Eya RNAi animals six days following amputation, 15 days following RNAi initiation. Data are mean ± s.e.m. (>12 animals per group). ***P<0.0001 determined by Student's t-test. Right: fluorescence in situ hybridization and immunolabeling of a regenerating control or Eya RNAi animal (tail blastema shown; anterior, left) using the CA (magenta) RNA probe and an anti-acetylated tubulin antibody (yellow). Scale bars: 0.05 mm. (B) Sall and Osr are expressed in tubule cells. Cartoon depicts blastema region imaged. Anterior, up. Scale bars: 0.05 mm. Insets on right show colocalization of transcription regulatory gene RNA (magenta) and cubilin (green). (C) Top: Sall(RNAi) animals bloated during tissue turnover in intact animals (17/20 animals bloated between days 43 and 46 after initial RNAi). Dotted box indicates the area depicted below. Middle: A bloated Sall(RNAi) animal with reduced tubule (Ac tubulin+, yellow) and increased tubule-associated cell numbers (CA+, magenta) is shown. Arrows indicate example clusters of ectopic tubule-associated cells at the animal periphery. Bottom: Higher magnification images of control and Sall(RNAi) clusters. Anterior, left. Scale bars: 0.05 mm.