Anti-HEXOKINASE (Yeast) (RABBIT) Antibody

Repression of HML and HMR preceded heterochromatin maturation.(A) Schematic of temperature-shift time course with sir3-8-M.ECOGII. (B) Protein immunoblotting in a strain expressing sir3-8-3xV5 (JRY13467) constitutively at 25°C (first lane), constitutively at 37°C (second lane), and at 30, 60, 90, 120, and 150 min after a shift to 25°C. Top row is 3xV5-tagged sir3-8 protein, the middle row is the same as the top row but at a higher exposure, and the bottom row is the loading control Hxk2. The unedited blot is in . (C) Aggregate methylation results at HML (top) and HMR (bottom) from long-read Nanopore sequencing of strains expressing sir3-8-M.ECOGII (JRY13114) grown constitutively at 25 or 37°C. Plots are as described in Figure 1D. (D) Aggregate methylation results at HML (top) and HMR (bottom) from long-read Nanopore sequencing of a strain expressing sir3-8-M.ECOGII (JRY13134) grown constitutively at 25°C (dotted gray line) and collected at 0, 15, 45, and 90 min after a temperature switch from 37 to 25°C. (E) RT-qPCR of HMLα2 (left) and HMRa1 (right) mRNA in strains expressing SIR3-M.ECOGII (black, JRY13027, JRY12840), sir3∆::M.ECOGII (green, JRY13029, JRY13030), or sir3-8-M.ECOGII (purple, JRY13114, JRY13134) collected at 0, 30, 60, 90, 120, and 150 min after a temperature switch from 37 to 25°C. Points are the average of three biological replicates and bars mark one standard deviation. DIP-seq of sir3-8-M.ECOGII (JRY13114).Shown are 10 kb regions centered at HML (left) and HMR (right). Cells were grown constitutively at either 25 or 37°C. Input results are plotted but not visible due to the strong DIP-seq signals. Nanopore sequencing over temperature switch time course (biological replicate).Aggregate methylation results at HML (top) and HMR (bottom) from long-read Nanopore sequencing of a strain expressing sir3-8-M.ECOGII (JRY13114) grown constitutively at 25°C (dotted gray line) and collected at 0, 15, 45, and 90 min after a temperature switch from 37 to 25°C. Figure provided by CiteAb. Source: Elife, PMID: 35073254.