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Fig. 3. Knockdown of CNK2 expression via RNAi. (A) Schematic representation of the CNK2-RNAi expression construct. The construct, designed to generate 834 bp of dsRNA upon expression, was used to produce stable RNAi strains by nuclear transformation. Bs, BsiWI; E, EcoRI; H, HindIII; N, NdeI; P, PstI restriction sites. (B) Northern analysis of CNK2 transcript levels in five independent CNK2-RNAi transformants. 15 µg total RNA was loaded in each lane. The blot was probed with 0.5 kb of 32P labeled CNK2 cDNA, stripped and reprobed with 0.5 kb of 32P labeled ß-tubulin cDNA. (C) The ratio of CNK2 message to tubulin message was determined by comparing band intensities using ImageQuant v2.0. CNK2-RNAi strain 20 was used in all subsequent experiments.





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