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. 2019 Mar 25;132(6):jcs226969. doi: 10.1242/jcs.226969

Fig. 3.

Fig. 3.

Mutations that cause loss of Set2-dependent H3K36 methylation are synthetic lethal with wee1-50. (A) WT, set2Δ, wee1-50 and set2Δ wee1-50 cells were serially diluted and spotted onto YES plates and incubated at the indicated temperatures for 2–3 days. (B) WT, set2-R255G, wee1-50 and set2-R255G wee1-50 cells were serially diluted and spotted onto YES plates and incubated at the indicated temperatures for 2–3 days. (C) H3ΔΔ (deletion of two of the three H3 genes), wee1-50, H3K36R, H3K36R wee1-50 cells were serially diluted and spotted onto YES plates and incubated at the indicated temperatures for 2-3 days. (D) Serial dilutions of WT cells overexpressing (O/E) empty vector pREP41x or pREP41x-JMJD2A (encoding hJMJD2A), and wee1-50 mutants expressing empty vector pREP41x or pREP41x-JMJD2A. Transformants were serially diluted and spotted onto EMM without leucine in the absence of thiamine at 25°C or 36°C. (E) Western blotting analysis of H3K9me3, H3K36me3, H3K36me2 and H3K36me1 in WT cells containing pREP41x or pREP41x-JMJD2A, and set2Δ cells. H3 is shown as a loading control. (F) Serial dilutions of WT cells overexpressing empty vector pREP41x or pREP41x-FBXL11, and wee1-50 mutants overexpressing empty vector pREP41x or pREP41x-FBXL11. Transformants were serially diluted and spotted onto EMM without leucine in the absence of thiamine at 25°C or 36°C. (G) Western blotting analysis of H3K36me3 and H3K36me2 in WT cells expressing pREP41x or pREP41x-FBXL11 and set2Δ cells. H3 is shown as a loading control.