Extended Data Fig. 5: Control experiment demonstrating the specificity of HaloTag staining in mRNA transport experiments. | Nature Neuroscience

Extended Data Fig. 5: Control experiment demonstrating the specificity of HaloTag staining in mRNA transport experiments.

From: Messenger RNA transport on lysosomal vesicles maintains axonal mitochondrial homeostasis and prevents axonal degeneration

Extended Data Fig. 5

WT i3Neurons co-expressing 3 HaloTags fused to PP7 coat protein and RPS7 fused to 24 PP7 RNA stem-loop repeats (top row) (see scheme in Fig. 4a), or untransfected WT i3Neurons (negative control) (bottom row), were grown on coverslips for 25 days and transduced with LAMP1-mNeonGreen (LAMP1-NG) for 36 h. Neurons were then incubated overnight with 200 pM of the fluorescent HaloTag substrate JF646 and imaged by spinning-disk confocal microscopy. Scale bars: 20 μm. Notice the absence of JF646 signal in the control i3Neurons, demonstrating that the JF646 signal represents RPS7 staining and not non-specific staining of lysosome-related vesicles or other vesicles. Experiments were repeated three times.

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