Fig. 2

eEF1Bγ co-localizes with specific mRNAs a The human Che-1 3′ UTR was folded according to the computer algorithm of Zuker and Stiegler to yield a structure of minimum free energy [65]. b Schematic representation of the chimeric bacteriophage MS2 coat protein fused to the GFP protein (MS2-GFP) and the reporter transcript containing multimers of the RNA stem-loop, recognized by the MS2-GFP protein, upstream of the 3’ UTR of the mRNA of interest named: “Report mRNA” (left panel). The MS2-GFP protein was expressed in HeLa cells either alone or with the Report mRNA carrying the 3′ UTR of Che-1 or vimentin mRNAs (upper panel). In the lower panel, MS2-GFP was co-expressed with myc-eEF1Bγ protein and with the Report mRNA carrying either the 3′ UTR of Che-1 or the 3′UTR of vimentin transcripts (right panel). c Co-localization of endogenous eEF1Bγ protein and either Che-1 or vimentin endogenous mRNAs in HeLa cells. Expression of eEF1Bγ was detected by indirect immunofluorescence using polyclonal eEF1Bγ antibodies (red), whereas Che-1 and vimentin mRNAs (green) were detected by RNA-FISH. Nuclei (blue) were stained with DAPI. Intensity correlation quotient (ICQ) shown in the bar graph was calculated using Coloc 2 plugin in the Image J/Fiji software and indicates whether the intensity of co-staining varies in synchrony over space. The values indicated represent an average over at least 10 cells from different images and the error bars indicate standard error