Figure 7
Par-1 and BRP are present within the same molecular complex. (A) Representative confocal stacks showing co-localization of Par-1 and BRP in the axons and boutons from third instar larvae of Par-1OE. Axons (Scale bar = 10 μm) and synaptic boutons (Scale bar = 5 μm) are stained with antibodies against BRP (Red), and Par-1 (Green). (B) Representative Western blot of proteins pulled down using the anti-BRP antibody and probed using anti-Par-1 antibody. Both the input and the IP were performed in the same blot and loaded on the same gel (different lanes). Beads only control shows no signal (C,D) Representative PLA signal (red) from larvae overexpressing Par-1 using GeneSwitch-ElavGal4 at T0, T24, T48, and T72 from axon bundles (C) and synaptic boutons (D). HRP marks presynaptic membrane boundary (blue) and arrows mark the punctate pattern and examples of brightest PLA signals. (E,F) Average PLA fluorescent intensity (A.U.) normalized to HRP intensity in axons bundles (E) and synaptic boutons (F). (n > 10). Error bars represent S.E.M. ****p ≤ 0.0001.
