ZFIN Image: Liao et al., 2019, Figure S5

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Figure Caption

Figure S5

ALS-Associated Mutations in ANXA11 Disrupt Its Interactions with Lysosomes, Related to Figure 5

(A) Co-localization of light-activated opto-ANXA11 or ANXA11 N-terminal domain or C-terminal domain with lysosomes in cells. U2OS cells were co-transfected with LAMP1-HaloTag, Opto-mcherry (CRY2olig-mcherry), Opto-ANXA11, Opto-ANXA11 NTD or Opto-ANXA11 CTD for 24 hrs. Cells with similar Opto-ANXA11 expression levels were exposed to 0.2% 488nm light to initiate oligomerization. Cells were imaged over 300 seconds of light activation. Scale bar: 2 μm.

(B) Percentages of light-activated Opto-mcherry (CRY2olig-mcherry), Opto-ANXA11 Opto-ANXA11 NTD or Opto-ANXA11 CTD clusters co-localizing with lysosomes after 300 seconds of light activation from the experiment in (A). n=14 (Opto-mcherry), 18 (ANXA11 full-length), 21(ANXA11 NTD), 21(ANXA11 CTD). One-way ANOVA, ns, not significant. ^∗∗∗∗p < 0.0001. Error bars = SEM.

(C) Frequency of LAMP1 labeled vesicles in axons expressed ANXA11 or ALS-associated ANXA11 mutants. n=25(WT), 50(D40G), 15(R235Q), 22(R346C). One-way ANOVA, ns, not significant. Error bars = SEM.

Acknowledgments

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Reprinted from Cell, 179, Liao, Y.C., Fernandopulle, M.S., Wang, G., Choi, H., Hao, L., Drerup, C.M., Patel, R., Qamar, S., Nixon-Abell, J., Shen, Y., Meadows, W., Vendruscolo, M., Knowles, T.P.J., Nelson, M., Czekalska, M.A., Musteikyte, G., Gachechiladze, M.A., Stephens, C.A., Pasolli, H.A., Forrest, L.R., St George-Hyslop, P., Lippincott-Schwartz, J., Ward, M.E., RNA Granules Hitchhike on Lysosomes for Long-Distance Transport, Using Annexin A11 as a Molecular Tether, 147-164.e20, Copyright (2019) with permission from Elsevier. Full text @ Cell