SCN2A, SCN3A and SCN9A are taste bud-associated genes. A-C, Isolation of mouse taste cells by LCM. CV papilla containing multiple taste bud areas before LCM (A), the same region after LCM (B), and the isolated taste bud areas (C). TB denotes taste bud and LE denotes lingual epithelium. LE adjacent to CV papilla and devoid of taste buds was also collected by LCM. Since tissue sections are not coverslipped and come into contact with an adhesive cap to remove collected regions during LCM, small lighting differences and focus changes are observed between images. Scale bar is 50 μm. D-F, RT-PCR amplification of transcripts from TB and LE collected by LCM. The taste bud-associated genes gustducin, T1R2, TRPM5, and PKD2L1 are expressed in TB but not LE, whereas the housekeeping genes β-actin and GAPDH are expressed in both TB and LE (D, F). SCN2A, SCN3A and SCN9A are taste bud-associated genes and are expressed in CV TB (E) and FG TB (F) but not LE. All primer sets amplified products from a pool of fifteen diverse tissues, denoted as 'Control' in the figure (Mouse Total RNA Master Panel; Clontech). Cloning and sequencing of PCR products confirmed that amplified bands corresponded to the expected genes. '+' indicates reverse transcription and '-' indicates no reverse transcription was performed. PCR products were only obtained following reverse transcription, indicating that bands originated from mRNA and not genomic DNA. G-K, ISH for TRPM5 (F), PKD2L1 (H), SCN3A (I), SCN2A (J) illustrating expression of transcripts in CV taste buds. Images in G-J depict antisense probes and image in K depicts a representative sense probe. No signals were observed with control sense probes. Scale bar in F is 50 μm and represents scale for G-K.