Subcellular localization of neuronal DCAF12. (A and B) Larval ventral nerve cord and eye disc stained for DCAF12 (GP11). (C) Neuronal somata stained for DCAF12 (GP12) and Lamin-C marking the nuclear envelope. (D) Larval NMJs stained for DCAF12 (GP11) and HRP marking the neuronal membrane. (E and F) Axons of NMJ (E) stained for DCAF12 and HRP and plot (F) of a single line scan (gray line in E) of DCAF12 and HRP fluorescence (n = 1). (G and H) Axons of NMJ stained for DCAF12 and Futsch (G) and plot (H) of a single line scan (gray line in G) of DCAF12 and Futsch fluorescence (n = 1). (I and J) Synaptic boutons stained for DCAF12 and postsynaptic DLG (I) or DCAF12 and HRP (J). (K) Frequency distribution of DCAF12 fluorescence in synaptic boutons of control (n = 118; n = 3) and Δ51 (n = 72; n = 3). (L) Synaptic boutons stained for DCAF12 and Brp (AZ). (M–O) Quantification of anti-DCAF12 fluorescence intensity (FI; means ± SEM; n ≥ 8; ***, P < 0.0002; two-tailed unpaired t test). Scale bars, 50 µm (A and B), 20 µm (D), 10 µm (E, I, and J), 5 µm (C and G), and 2.5 µm (L).