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Fig. 6. Knockdown of Robo family proteins using transgenic RNAi. (A) Western blot analysis of adult heads, showing that animals homozygous for robo3¹ express full-length Robo3 protein (caret) as well as a truncated Robo3 (asterisk). Anti-Elav used as loading control. (B) Western blot analysis of adult heads, showing that expression of UAS:RoboRNAi in the nervous system controlled by c155-Gal4 reduces Robo protein levels. (C) Ubiquitous expression of UAS:Robo2RNAi under the control of tubulin-Gal4 reduces anti-Robo2 staining in the visual system. (D) Western blot analysis of adult heads, showing that expression of UAS-Robo3RNAi controlled by c155-Gal4 reduces Robo3 protein levels. (E-M) RNAi of an individual Robo family member does not detectably reduce expression of other Robo family proteins. (E-G) Anti-Robo staining in magenta. (H-J) Anti-Robo2 expression in magenta. (K-M) Anti-Robo3 expression in magenta. (E,H,K) Robo RNAi detectably reduces Robo expression (open arrowhead), but not Robo2 or Robo3 (closed arrowheads). (F,I,L) Robo2 RNAi detectably reduces Robo2 expression (open arrowhead), but not Robo or Robo3 (closed arrowheads). (G,J,M) Robo3 RNAi detectably reduces Robo3 expression (open arrowhead), but not Robo or Robo2 (closed arrowheads).

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