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. 2016 May 18;12(5):e1006041. doi: 10.1371/journal.pgen.1006041

Fig 6. Ppk32 function is regulated by conserved phosphorylation.

Fig 6

All strains are leucine autotrophs. (A). Sequence alignment between Ppk32 and other SCYL family kinases. (B) Early exponential cells were grown in rich medium (YES). Samples from non-stressed cultures were taken for Western blot analysis of Ppk32 level. Ponceau S staining was used as a loading control, * indicates a background band. (C) Growth assay. Exponentially YES-grown cells of indicated strains were spotted in 10-fold serial dilution onto the indicated media. (D) Tor1 immuno-precipitations first 3 lanes (Fig 1) are shown again for comparison. Soluble proteins were extracted from exponentially grown strains. Tor1 immuno-precipites and Ppk32 co-immuno-precipites were assessed from the same gels. The relative levels of specific Ppk32 immuno-precipitation are shown. (E-F) Early exponential indicated strains were grown in minimal media (EMMG) to achieve Ppk32 over-expression. Shifted from 28°C to 37°C to induce heat stressed (F) Western blot analysis of phosphorylated S630 and S632 of Ppk32 and total Ppk32 levels. (G) Quantification of relative Ppk32 S630 and S632 phosphoryaliton after 4 hours heat stress.