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. 2018 Mar 12;14(3):e1007253. doi: 10.1371/journal.pgen.1007253

Fig 9. Expression of SUT075 in trans rescues the lethal phenotype of a SUT075 deletion and increases PRP3 expression.

Fig 9

(A) Haploid spores from dissection of six MATa/α SUT075Δ/SUT075 diploid tetrads were spotted on SD media lacking uracil and containing 2% galactose. The plasmid expressing the SUT075 ncRNA is selected for using the URA auxotrophic marker. Galactose induces expression of SUT075 present in the plasmid. (B) Haploid spores from dissection of six MATa/α SUT075Δ/SUT075 diploid tetrads were spotted on SD media lacking uracil containing 2% galactose and 300mg/L G418 disulphate. G418 resistance selects for haploids deleted for SUT075. Spores growing in both panels A and B are considered to contain the G418 resistance SUT075 deletion cassette and the SUT075 ncRNA expressing plasmid. (C) Expression levels of PRP3 in the ΔSUT075 and the ΔSUT075 (+ sense SUT075 recovery plasmid) heterozygote diploid strains measured by qRT-PCR. The relative expression of PRP3 in the wild-type background is represented by a grey bar and a black bar in the ncRNA deletion strain backgrounds. Using the ΔΔCт method and ACT1 as a reference gene, the fold change (2^) in expression, relative to the wild-type was calculated. Error bars are calculated using each of the three independent biological samples. P values calculated using the Welch two sample t-test determine there to be a significant difference (p = 0.02) between the ΔSUT075 and the ΔSUT075 (+ sense SUT075 recovery plasmid) strains.