Fig. 2.

Single-cell RT-PCR of red-fluorescent cells from Trpc2-IRES-taumCherry mice. (A) Ethidium-bromide stained agarose gels of RT-PCR products generated from a single red-fluorescent cell (m37) isolated from lateral WOM of a homozygous Trpc2-IRES-taumCherry mouse at eight weeks, in comparison with RT-PCR products of WOM of a WT C57BL/6 mouse at eight weeks (OE), PCR products of genomic DNA (mut) from a homozygous Trpc2-IRES-taumCherry mouse as positive control for mCherry, and buffer (buf) as negative control. (B) Degenerate RT-PCR with primers for OR genes does not reveal OR gene expression in 15 single cells isolated from lateral WOM of a homozygous Trpc2-IRES-taumCherry mouse at eight weeks. We cloned some of the faint bands and found that they did not represent OR genes. A strong RT-PCR product is obtained with o1 and o2, two single cells isolated from WOM of a heterozygous OMP-YFP gene-targeted mouse at three weeks. buf16 is buffer, as negative control. (C) Degenerate RT-PCR for the V1rg family of Vmn1r genes reveals a product in one of six single red-fluorescent cells (v10) isolated from the VNO a homozygous Trpc2-IRES-taumCherry mouse at eight weeks. By contrast, no RT-PCR products are obtained from nine single red-fluorescent cells (such as m2) picked from lateral WOM. Lanes labeled with buf are with buffer, as negative control. (D) RT-PCR for the family-C Vmn2r genes reveals a product in a single red-fluorescent cell (v28) isolated from the VNO a homozygous Trpc2-IRES-taumCherry mouse at eight weeks, but not from six single red-fluorescent cells (such as m62) isolated from lateral WOM of a homozygous Trpc2-IRES-taumCherry mouse at eight weeks. A and B are two distinct primer sets. Lanes labeled with buf are with buffer, as negative control. VNO represents cDNA from whole vomeronasal organ, and Gen represents genomic DNA of a WT C57BL/6 mouse.