<p>A. Pull-down assay between GST-tagged dPur α and <i>in vitro</i>-translated Rm62 or <i>in vitro</i>-translated Hts or luciferase (negative control). Indicated are samples treated or untreated with RNase prior to binding reaction. In the Input lanes, we loaded 25% of the translation products used in a reaction. Both Rm62 and Hts, but not luciferase, interacts with dPur α. B. Western blot shows the interaction between endogenous Rm62 or endogenous Hts and affinity-purified GST-Pur α. Indicated are samples treated or untreated with RNase prior to binding reaction. C. Western blot shows the interaction between affinity-purified GST-Rm62 and endogenous Pur α in RNase untreated and treated samples. D. Western blot shows the interaction between endogenous mammalian p68 and affinity-purified fly GST-Pur α. E. Protein levels of Rm62 and Hts in wild-type and rCGG-expressing flies. Quantitative analysis of Rm62 and Hts protein levels by Western blot on adult head extracts of the following genotypes: wild-type (WT); <i>elav</i>; rCGG<sub>60</sub>-expressing flies. Proteins are indicated to the right, corresponding molecular weights to the left. α actin represents a loading control. F. Quantitative analysis of <i>Rm62</i> mRNA levels by real-time PCR on total RNA obtained from adult heads of wild-type (WT) and rCGG-expressing flies. Quantification is relative to the housekeeping ribosomal protein 32 (<i>Rpl32</i>) mRNA. (E; mean ± SEM n = 3). G. Statistical evaluation of the percent viability displayed by various genotypes: <i>elav/+; +/+; Rm62<sup>LOF</sup>/+</i> (Rm62 heterozygous); <i>elav/+; rCGG<sub>60</sub>-EGFP/+; TM3Sb/+</i> (Premutation heterozygous), <i>elav/+; +/+; TM3Sb/+</i> (Internal control); <i>elav/+; rCGG<sub>60</sub>-EGFP/+; Rm62<sup>LOF</sup>/+</i> (interaction). Mean of three data sets was used.</p
