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. 2014 Nov 20;10(11):e1004783. doi: 10.1371/journal.pgen.1004783

Figure 5. HAM-5 is required to localize MAK-2 and MEK-2 to puncta.

Figure 5

(A) MAK-2-GFP in isolated WT germlings localizes to the nucleus, cytoplasm and to puncta (white arrows). (B) In the Δham-5 mutant, MAK-2-GFP localization is cytoplasmic and nuclear; no puncta are observed. Scale bar  = 10 µM. (C) In WT hyphae, MEK-2-mCherry localizes to the septum (green arrow) and also to cytoplasmic puncta (white arrow). Scale bar  = 10 µM. (D) In Δham-5 hyphae, septum localization of MEK-2-mCherry is observed (green arrow), but puncta are not. Scale bar  = 10 µM. (E) Co-immunoprecipitation experiments showing an interaction between MEK-2-mCherry and phosphorylated MAK-2 in WT (mek-2-mCherry) germlings, but a significant reduction in interaction between MAK-2 and MEK-2-mCherry in a Δham-5 (mek-2-mCherry) strain. Top panel is a Western blot of protein samples probed with anti-p42/44 antibodies (MAK-2, 40.8 kD, MAK-1, 46.7 kD; Figure S4E). Middle panel is anti-mCherry immunoprecipitated proteins probed with anti-mCherry antibodies (MEK-2-mCherry, 82.9 kD; Figure S4C, E). Bottom panel is anti-mCherry (MEK-2-mCherry) immunoprecipitated proteins probed via Western blot with anti-p42/44 antibodies that recognize phosphorylated MAK-2.