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. 2008 Mar 5;28(10):2332–2341. doi: 10.1523/JNEUROSCI.4807-07.2008

Figure 2.

Figure 2.

Two- and three-color ISH reveal three layers of VSNs. A, Two-color ISH with the mixed H2-Mv probe (red) and the GFP probe (green) on a VNO section of a V2r1b–IRES–tauGFP mouse shows nonoverlapping expression. B, Two-color ISH with the mixed H2-Mv probe (red) and the V2rf4-specific probe (green) shows nonoverlapping expression. C, D, Three-color ISH with the mixed H2-Mv mixed probe (red), the Gαi2 probe (green), and the V2r1b probe (blue) (C) or the V2rf1–3 probe (blue) (D). Overlay of red and blue results in purple. E, Three-color ISH with the V1rb1 probe (red), the V2rf1–3 probe (green), and the V2r1b probe (blue). F, The apical–basal positions of V1rb1-, V2rf1–3-, and V2r1b-labeled cell bodies in E were measured, with 0 for basal and 1 for apical. They are distributed in distinct layers: V1rb1, 0.52 ± 0.12 (n = 89); V2r1b, 0.26 ± 0.13 (n = 196); V2rf1–3, 0.15 ± 0.11 (n = 159). *p < 0.01, **p < 0.01. G, H, Three-color ISH with the Gαi2 probe (green), the V2r1b probe (blue), and the Gαo probe (red) (G) or the β2m probe (red). H shows overlap of V2r1b signals with both Gαo and β2-microglobulin. I, Three-color ISH with the mixed H2-Mv mixed probe (green), the Mhc-pan probe from H2-Q1 (red), and the V2r1b probe. V2r1b+ neurons are also Mhc-pan−. Arrowheads point to V2r1b+ cell bodies in C and G–I and to V2rf1–3+ cell bodies in D. All samples were from 8- to 10-week-old male mice. Wild-type mice were used in B–I. Scale bars, 50 μm.