(a) Quadruple immunohistochemistry for ARTN, TH, ASMACy3 and CD31 on P3 and P11 NEM sections from wild type mice. TH+ sympathetic axons were observed associated with blood vessels (arrowheads) and at the base of the NEM at P3 and on the ARTN-expressing NEM at P11 (arrows). (b) Triple immunohistochemistry for NRTN, TH and ASMACy3 on P5 and P7 PEM sections from wild type mice. Note TH+ sympathetic axons (arrowheads) at the base of the PEM at P5 and on NRTN expressing PEM at P7 (arrow). (c) Quadruple immunohistochemistry for ASMACy3, ARTN, CD31 and DAPI on P5 NEM sections from wild-type mice. CD31+ASMACy3+ blood vessel cells (arrows) in close association with the NEM express lower levels of ARTN, when compared to ASMACy3+ NEM cells (arrowheads). (d) Quadruple immunohistochemistry for ASMACy3, ARTN, K5 and DAPI on adult NEM sections from wild type mice. ASMACy3+ blood vessel cells (dashed lines) in close association with the NEM express little or no ARTN. (a-d) are representative images from experiments carried out on at least two animals. Scale bars: 10 μm in all images.