Fig. 2Components of the endosomal mRNA transport. (a) DIC image of an Rrm4-GFP expressing hypha 6 h
after induction of hyphal growth with corresponding fluorescence image and kymograph (scale bar, 10μm,
inverted fluorescence image). (b) Example for velocity measurement within a kymograph generated by the
software package MetaMorph (inverted image). Thered lineindicates a processively moving signal from the
hyphal tip toward the basal pole (righttoleft). Based on the position, distance and elapsed time the velocity
was calculated. (c) Schematic representation of the components of the RNA live imaging technique inU.
maydis. Theupper partshows the RNA-binding protein (RBP) fused to three fluorescence proteins (FP)
expressed at the ectopicipslocus inU. maydis. Expression is either driven by an arabinose-inducible promoter
Pcrgor a constitutively active promoter Potef. Thelower partshows the locus of the gene of interest (GOI). Its
expression is either driven by Potefor by the endogenous promoter. 16 boxB stem-loops (red) are inserted in
the 3^0 UTR. (d) Micrograph showingcdc3mRNA signals in hypha indicated byblack arrowheads(top, scale bar
10 μm, inverted image). Kymograph visualizes movement ofcdc3mRNA (black arrowheads). (e) Kymographs
of the described strain to visualize the colocalization of mRNA (green) and Rrm4 (red).Arrowheadmarks
colocalization event (inverted images). (f) Kymographs of the described strain to visualize the colocalization of
mRNA (green) and encoded protein (red).Arrowheadsmark colocalization event (inverted images)
RNA Live Imaging inU. maydis 327