Fig. 3. Differential expression of Slit proteins in the AOB. The experiments were carried out on sagittal sections of the AOB at P1. (A) Schematic drawing of the probes used. As Ig domains 1 and 2 confer Slit binding, a deletion construct omitting these two domains named (1,2)-Robo1-Fc was used as a control, while (3,4,5)-Robo2-Fc containing Ig domains 1 and 2 was used to detect Slit protein expression. (B) Dorsal view of the AOB. Basal vomeronasal axons (green) invade the AOB from its medial margin and project onto the posterior AOB. Approximate location of sagittal sections shown in C,D,E are indicated by broken lines i, ii and iii, respectively. (C,D,E) Detection of Slit proteins using (3,4,5) Robo1-Fc on 150 µm sagittal vibratome sections. Their approximate positions along the mediolateral axis (i,ii,iii) are shown in B. Bound (3,4,5) Robo1-Fc was visualised using an alkaline phosphatase conjugated anti-Fc specific antibody. (G-I) Corresponding DAPI stainings to C,D,E. (F,J) Staining with (1,2) Robo2-Fc (F) serving as a negative control and the corresponding DAPI staining (J). (K-N) Quantification of staining patterns shown in C-F using the Phoretix ID Quantifier V4.0 program. Here, the staining intensity of the area between the two asterisks shown in C-F, which covers the AOB along its entire anteroposterior dimension, was measured. Graphs show average values and standard deviations. K shows the results from the measurement of six comparable sections (n=6) at the level of i, of which C is a typical example (n=10 for L, n=6 for M and n=3 for N). These data indicate a stronger expression of Slits in the anterior than the posterior AOB. AOB, accessory olfactory bulb; A, anterior; D, dorsal; L, lateral; M, medial; MOB, main olfactory bulb; P, posterior; V, ventral. Scale-bar: 100 µm in C-J.

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