Extended Data Fig. 7: BORC-independent rescue of axonal RPS27A mRNA by expression of LAMP1-3xKBS in BORCS5-KO and BORCS7-KO i3Neurons. | Nature Neuroscience

Extended Data Fig. 7: BORC-independent rescue of axonal RPS27A mRNA by expression of LAMP1-3xKBS in BORCS5-KO and BORCS7-KO i3Neurons.

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

Extended Data Fig. 7: BORC-independent rescue of axonal RPS27A mRNA by expression of LAMP1-3xKBS in BORCS5-KO and BORCS7-KO i3Neurons.

a, WT, BORCS5-KO and BORCS7-KO i3Neurons co-expressing the PP7 coat protein fused with three HaloTags and RPS27A CDS fused with 24 PP7 RNA stem-loop repeats were grown on coverslips for 15 days. Cells were then transduced with a construct encoding LAMP1-3xKBS-NG for 36 h and incubated with 200 pM of the fluorescent Halo substrate JF646 overnight to image lysosome-related vesicles and RPS27A mRNA localization at axon terminals. Scale bars for left, LAMP1-3xKBS-NG and merged columns: 10 μm. Scale bars for RPS27A zoomed-in boxed areas: 5 μm. b, Quantification of the number of RPS27A puncta per axon terminal from n = 3 independent experiments. Values are represented as SuperPlots, showing the individual data points and the mean of the means ± SD. Statistical significance was calculated by one-way ANOVA with Dunnett’s multiple comparisons test. BORCS5 KO vs WT ***P < 0.001, BORCS7 KO vs WT ***P < 0.001, BORCS5 KO + LAMP1-3xKBS vs BORCS5 KO ***P < 0.001, BORCS7 KO + LAMP1-3xKBS vs BORCS7 KO ***P < 0.001, BORCS5 KO + LAMP1-3xKBS vs WT + LAMP1-3xKBS P = 0.721, BORCS7 KO + LAMP1-3xKBS vs WT + LAMP1-3xKBS P = 0.136. ns: not significant relative to WT. Notice the increased number of RPS27A mRNA and lysosomal puncta in BORCS5-KO and BORCS7-KO i3Neurons transduced with LAMP1-3xKBS-NG.

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