Fig 4. MLLE3 is key for endosomal mRNA transport.

(A) Schematic representation of Rrm4 and Upa1 variants drawn not to scale (number of amino acids indicated next to protein bars) using the following colouring: dark green, RNA recognition motif (RRM); orange, MLLE domains; red, mKate2, blue, PAM2, light orange PAM2 like sequence (PL1–2) and light green, Gfp. Ankyrin repeats (5xANK), FYVE domain and RING domain of Upa1 are given in dark grey. (B) Growth of AB33 derivatives in their hyphal form (6 h.p.i.; size bar 10 μm). Growth direction is marked by arrows. (C) Quantification of hyphal growth of AB33 derivatives shown in B (6 h.p.i.): unipolarity, bipolarity and basal septum formation were quantified (error bars, SEM.; n = 3 independent experiments, > 100 hyphae were counted per strain; for statistical evaluation, the percentage of uni- and bipolarity was investigated by using unpaired two-tailed Student’s t-test (α<0.05). (D) Kymographs of AB33 hyphae derivatives (6 h.p.i.; inverted fluorescence images) expressing pairs of red and green fluorescent proteins as indicated. Fluorescence signals were detected simultaneously using dual-view technology (arrow length on the left and bottom indicates time and distance, respectively). Processive co-localising signals are marked by red arrowheads. (E) Percentage of processive signals exhibiting co-localisation for strains shown in D (data points represent means from n = 3 independent experiments, with mean of means, red line and SEM; unpaired two-tailed Student’s t-test (α<0.05); for each experiment, 10 hyphae per strains were analysed).