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. 2008 May 19;28(14):4445–4458. doi: 10.1128/MCB.00164-08

FIG. 1.

FIG. 1.

gcn5 mutants lacking either Rpd3(L) or Rpd3(S) are sick. (A) Tenfold dilutions of strains DY5699 (wild type), DY10398 (rco1), DY11148 (sds3), DY10460 (rco1 sds3), DY11354 (gcn5), DY11355 (gcn5 rco1), DY11357 (gcn5 sds3), and DY11359 (gcn5 rco1 sds3) were plated on complete medium for 3 days at either 25°C or 30°C. (B) Tenfold dilutions of strains DY5699 (wild type), DY4895 (rpd3), DY11354 (gcn5), DY5169 (gcn5 rpd3), and DY11359 (gcn5 rco1 sds3) were plated on complete medium for 2 days at either 25°C or 30°C. (C) Strains DY150 (wild type) and DY11087 (gcn5) were transformed with the indicated multicopy plasmid, and 10 dilutions were plated on selective medium lacking uracil for 3 days at 30°C or for 3 days at 37°C. (D) Strains DY13050 (Sin3-HA Rco1-Myc Sds3-Myc), DY13056 (Sin3-HA rco1Δ Sds3-Myc), and DY13082 (Sin3-HA Rco1-Myc sds3Δ) were grown in synthetic complete medium and strain DY13050 (Sin3-HA Rco1-Myc Sds3-Myc) transformed with plasmid M5340 (YEp-RCO1-Myc) was grown in selective medium lacking uracil. Extracts were prepared and immunoprecipitated with anti-HA antibody (anti-HA IP) or with no antibody as a control (no Ab IP). The immunoprecipitated proteins were electrophoresed and transferred to Western blots, along with a control corresponding to 5% of the input before immunoprecipitation (5% input), and the blots were probed with anti-HA and anti-Myc antibodies. Protein bands in the immunoblots were quantitated, with Rpd3(S) levels corresponding to the ratio of Rco1-Myc to Sin3-HA and Rpd3(L) levels corresponding to the ratio of Sds3-Myc to Sin3-HA. (E) Tenfold dilutions of strains DY5699 (wild type), DY10398 (rco1), DY11148 (sds3), and DY10460 (rco1 sds3) were plated on complete medium for 3 days at 30°C or on medium containing 100 mM HU for 5 days at 30°C. (F) Tenfold dilutions of strains DY5699 (wild type), DY10398 (rco1), DY8825 (set2), DY10402 (set2 rco1), DY11087 (gcn5), DY11504 (gcn5 rco1), DY11509 (gcn5 set2), and DY11505 (gcn5 rco1 set2) were plated on complete medium for 3 days at 25°C or for 2 days at 30°C. (G) Tenfold dilutions of strains DY5699 (wild type), DY11148 (sds3), DY8825 (set2), DY10430 (set2 sds3), DY11087 (gcn5), DY11357 (gcn5 sds3), DY11509 (gcn5 set2), and DY11513 (gcn5 sds3 set2) were plated on complete medium for 4 days at either 25°C or 30°C.