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. 2015 Apr 24;11(4):e1005197. doi: 10.1371/journal.pgen.1005197

Fig 7. Effects of K63 polyubiquitination and Hel2 on quality control for non-stop translation.

Fig 7

(A) Genetic colony growth test for the nonstop HIS3 reporter. A schematic image of a nonstop HIS3 allele is illustrated above. Empty vector (empty), wild-type ubiquitin (Ubi-WT), or K63R ubiquitin (K63R) plasmids were introduced into the wild-type, ski3 ∆, hel2 ∆, or ltn1 ∆ strains harboring nonstop HIS3 reporter (S17-A08, S17-A09, SKY123, SKY137). Types of knockout alleles and vectors are indicated on the left. Transformant colonies were streaked on the SC-Uracil (Ura) (middle), and SC-Uracil, Histidine (Ura, His) (right) plates, and colony growth was monitored for 3 days at 30°C. (B) Western blot analysis for the nonstop HIS3 reporter. Protein samples were extracted from the transformants of the nonstop HIS3 reporter strains as in (A). The expression of HIS3 protein was detected using a HIS3 polyclonal antibody. PGK1 was used as a loading control. (C) Northern blot analysis for the nonstop HIS3 reporter. mRNA samples were extracted from the transformants of the nonstop HIS3 reporter strains as in (A). The nonstop HIS3 reporter mRNA was probed by HIS3 complementary DNA fragment. SCR1 mRNA was detected as a loading control.