Fig. 2.

Calorie restriction does not affect subtelomeric gene silencing. (A) Qualitative silencing assay. The telomeric silencing reporter parent strain, AEY1017, and its isogenic sir2Δ strain were grown up in YPD, then diluted in H₂O and serially spotted out on synthetic complete media (SC) and SC + FOA media (FOA). Spots correspond to OD₆₀₀ = 1 (left) then serial 1 in 5 dilutions to the right. Plates were imaged after 2 (0.5% and 2% glucose) and 3 days (0.05% glucose) at 30 °C. Reduced growth on FOA compared with SC indicates reduced silencing. (B) Quantitative silencing assay. Serial dilutions of overnight cultures were spread on plates and colonies were counted after 2 (0.5% and 2% glucose) or 3 days (0.05% glucose). Silencing is proportional to viability on FOA media, which was calculated by dividing the numbers of colonies on FOA plates by the total numbers of colonies on −ura and FOA plates combined. 62 000 colonies were counted in total. The graph shows pooled data normalized to the 2% glucose control value, expressed as mean + standard error of the mean. No statistically significant difference between the conditions was found.