Specification of vulval precursor cell (VPC) fates in C. elegans has served as an important signal transduction paradigm. Genetic studies have indicated that a large group of synthetic multivulva (SynMuv) genes, including the Rb ortholog lin-35, antagonizes the activity of the EGF receptor-Ras-MAP kinase pathway during VPC specification. A prevalent view has been that Rb-mediated transcriptional regulation and chromatin remodeling activities act in the VPCs to antagonize Ras activation through effects on promoters of target genes of the EGF receptor-Ras-MAP kinase pathway that promote vulval fates. Here, we have investigated the cellular focus of lin-35 using conventional genetic mosaic analysis and tissue-specific expression. Our results indicate that lin-35 activity is required in the major hypodermal syncytium and not in the VPCs to inhibit vulval fates. LIN-35 Rb may inhibit vulval fates by regulating a signal from hyp7 to the VPCs or the physiological state of hyp7.