Extended Data Fig. 7: Heritable silencing of a KanR-ade6⁺ transgene inserted at four different genomic loci.

a–d, Schematic diagrams of KanR-ade6⁺ insertions at the mal1⁺ (a), efm3⁺ (b), meu10⁺ (c), and mrp1⁺ (d) loci. e–h, KanR-ade6⁺ OFF cells of the indicated genotypes were crossed to cen1⁺ ade6-M210 cells and RSA was performed. The number of progeny matching each genotype and phenotype are shown. Total red or white colonies genotyped are indicated for each cross. For red cells, the first number indicates kanamycin-resistant colonies (indicating presence of the KanR-ade6⁺ OFF transgene) and second number indicates total red colonies (the remainder of which possess the endogenous ade6-M210 mutant allele). i, cen⁺ KanR-ade6⁺ mlo3⁺ leo⁺ progeny of the crosses in e–h were plated on media containing low adenine to test for stability of each epiallele during mitotic growth. Experiments were performed once. j, ChIP–qPCR assay showing enrichment of H3K9me3 at KanR-ade6⁺ epialleles in the cen⁺ mlo3⁺ leo1⁺ progeny of the crosses in e–h. Data are mean ± s.d. from three biological replicates. k, l, efm3::KanR-ade6⁺ OFF (k) or meu10::KanR-ade6⁺ OFF (l) cells were crossed to ade6-M216 ago1∆ (left) or clr4∆ (right) and RSA was performed. All ade6⁺ OFF progeny were ago1⁺ and clr4⁺. Bars indicate the number of ade6⁺ OFF meiotic progeny for each genotype. Total red or white colonies genotyped are indicated for each cross. For red cells, the first number indicates kanamycin-resistant colonies (reflecting the presence of the KanR-ade6⁺ OFF transgene) and the second number indicates the total red colonies (the remainder of which possess the ade6-M210 allele).