Figure 2
FRUMis significantly reduced in En-fruMRNAi males. CNSs of wild-type (A, C, C’ E-G) and En-fruMRNAi (B, D, D’, H-J) males labeled for anti-FRUM (green) and anti-En (magenta) expression. (A) FRUM expression in E/F-brain neurons. (B) FRUM is extremely reduced in the E/F-brain neurons of En-fruMRNAi males. In the brain, FRUM is reduced by 86% (n = 3, compare A to B, see results for methods). (C) FRUM expression in E/F-VNCmid T1 (arrow) and E/F-VNCmed T1 and T2 (arrowheads) neurons. (D) FRUM is reduced by 81% in midline (n = 3) and by 77% in medial (n = 3, arrowheads) neurons in En-fruMRNAi males. (E-G) FRUM expression in ventral (E), ventromedial (F) and medial (G) AbG neurons (arrows). (H-I) FRUM is reduced by 77% in AbG neurons (n = 3) of En-fruMRNAi males. To clearly show that FRUM has been reduced in all AbG neurons, panels E, F, and G are at three different z-axis positions of the AbG in a WT male and panels H, I, and J are at similar z-axis positions in an En-fruMRNAi male. Arrowheads in panels H-J denote neurons that appear to co-express En/FRUM (but do not) because of overlap of two separate neurons in adjacent z-sections. (C’-D’) Examples of FRUM-only reference neurons (arrowheads) and En/FRUM neurons (arrows) are shown in panels C’ (WT male) and D’ (En-fruMRNAi male). Images are either confocal z-stacks (A-D, E-J) or overlays of single z-sections (C’, D’). Size bars = 20 μm (A) for panels A-D, and (E) for panels E-J.