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. 2013 Dec 26;9(12):e1004071. doi: 10.1371/journal.pgen.1004071

Figure 8. Smc5/6-depleted cells progress relatively normally through meiotic prophase I and enter nuclear divisions with damaged DNA.

Figure 8

(A) Western blot analysis of Mec1 substrates, Hop1 (pT318) and H2A (pS129, γH2A). Clb1 and Clb3 are meiosis I- and meiosis II-specific B-type cyclins, respectively [101]. Pgk1 is a loading control. Strains: WT (Y4567), smc5 (Y4570), and nse4 (Y4573). Spindle pole body separation was used as a marker of cell cycle progression. (B) Western blot of Rec8-GFP and Pds1-13Myc. Strains: WT (Y2572), smc5 (Y2673), and nse4 (Y3653). (C) Typical examples of immunofluorescence images of γH2A at anaphase I from wild type and the two mutants. Bars: 2 µm. Right: quantification of the number of γH2A foci directly localized to the DNA. WT (Y1381), smc5 (Y2704), and nse4 (Y2705). (D) Typical examples of immunofluorescence images from wild type and smc5 undergoing meiosis II. Bars: 2 µm. (E) Sporulation frequencies at 24 hours in smc5 and nse4 mutants in combination with mutants that show robust prophase I arrest. Strains: WT (Y1381), smc5 (Y2704), and nse4 (Y2705), dmc1 (Y2045), dmc1 smc5 (Y3491), dmc1 nse4 (Y3488), dmc1 nse4 fpr3 (Y4606), rec8 (Y4607), rec8 smc5 (Y2856), rec8 nse4 (Y2855), hop2 (Y2489), hop2 smc5 (Y4610), hop2 nse4 (Y4613), hop2 nse4 fpr3 (Y4616).