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. 2020 Sep 1;48(17):9822–9839. doi: 10.1093/nar/gkaa699

Figure 4.

Figure 4.

Task3 5′ UTR (GGN)13 repeat is required for distal neurite localisation of Task3 mRNA. (A) RNA FISH of DIV10 embryonic murine cortical neurons transfected with WT G4 Task3M1-eGFP. Cells were hybridised with anti-eGFP probes and anti-MAP2/Tau primary antibodies to determine neurite localisation of exogenous Task3 mRNA. Image 2 was taken at two exposures to define mRNA granules in both the cell body and neurites with the respective images merged (dotted line). Original un-edited images available in Supplementary Figure S5. (B) RNA FISH of transfected DIV10 embryonic murine cortical neurons co-transfected with mutant G4 5′ UTR Task3M1-eGFP and Fluc-3F as a soluble whole-cell marker. Cells were hybridised with anti-eGFP probes and anti-FLAG M2 primary antibody showing localisation of exogenous mutant G4 5′ UTR Task3M1-eGFP mRNA in comparison to whole-cell staining. (C) The distance of exogenous mutant G4 5′ UTR Task3M1-eGFP mRNA from the cell body, quantified in Fiji for cells possessing neurites ≥100 μm in length. Number of neurites analysed is shown from three blinded biological replicates (coloured groups), where error bars represent SEM. Statistical analyses were carried out by one-way ANOVA (F(3, 247) = 89.01, P < 0.0001), with post-hoc Sidak's multiple comparison tests, P values shown. Scale bar = 20 μm.