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. 2019 Dec 2;8:e49494. doi: 10.7554/eLife.49494

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© 2019, Armijo-Weingart et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 3. — (A) Timelapse sequence showing the formation of an actin filament patch at the site of fission during the first 10 min of NGF treatment. The initiation of the actin patch is shown at time 0 s (green arrow) and tracked throughout the process of fission. The patch elaborates at the future site of fission (red arrow) that becomes evident at 96 s. Other actin patches formed along the mitochondrion are not associated with sites of fission. (B) Pretreatment (15 min) with the actin depolymerizing drug Latrunculin A (LatA; 4 μM) prior to NGF treatment (0–10 min) blocks the NGF-induced increase in the rate of mitochondria undergoing fission (% mitochondria/10 min). Each data point reflects one axon. Dunn’s posthoc multiple comparison tests. NGF and no NGF treatments included treatment with the DMSO vehicle. Median is denoted by arrowheads to the right of data points. (C) LatA pretreatment blocks the NGF-induced increase in the formation of Drp1 accumulations along mitochondria. Same experimental design as in (B), n = mitochondria(axons) shown in bars. Dunn’s posthoc multiple comparison tests. (D) Example of actin patch formation associated with a mitochondrion that gives rise to the emergence of a filopodium but not fission. By 4 s an actin patch has begun forming at the site populated by the mitochondrion (yellow bracket at 4–30 s). At 30 s a filopodium emerges from the patch (arrow). The heat map ranges 0–4095 in pixel intensities. (E) Analysis of the duration of actin patches during the first 10 min of NGF treatment as a function of their relationship to mitochondria and sites of fission. n = number of patches from 19 axons. Dunn’s posthoc multiple comparison tests. (F) Analysis of the probability that an actin patch will give rise to a filopodium as a function of their relationship to mitochondria and sites of fission. Same data set as in (E). Fisher’s exact tests.

Figure 3—figure supplement 1. — (A) Timelapse sequence showing the formation of an actin filament patch at the site of fission during the first 10 min of NGF treatment. The initiation of the actin patch is shown at time 0 s (green arrow) and tracked throughout the process of fission. The patch elaborates at the future site of fission (red arrow) that becomes evident at 96 s. Other actin patches formed along the mitochondrion are not associated with sites of fission. (B) Pretreatment (15 min) with the actin depolymerizing drug Latrunculin A (LatA; 4 μM) prior to NGF treatment (0–10 min) blocks the NGF-induced increase in the rate of mitochondria undergoing fission (% mitochondria/10 min). Each data point reflects one axon. Dunn’s posthoc multiple comparison tests. NGF and no NGF treatments included treatment with the DMSO vehicle. Median is denoted by arrowheads to the right of data points. (C) LatA pretreatment blocks the NGF-induced increase in the formation of Drp1 accumulations along mitochondria. Same experimental design as in (B), n = mitochondria(axons) shown in bars. Dunn’s posthoc multiple comparison tests. (D) Example of actin patch formation associated with a mitochondrion that gives rise to the emergence of a filopodium but not fission. By 4 s an actin patch has begun forming at the site populated by the mitochondrion (yellow bracket at 4–30 s). At 30 s a filopodium emerges from the patch (arrow). The heat map ranges 0–4095 in pixel intensities. (E) Analysis of the duration of actin patches during the first 10 min of NGF treatment as a function of their relationship to mitochondria and sites of fission. n = number of patches from 19 axons. Dunn’s posthoc multiple comparison tests. (F) Analysis of the probability that an actin patch will give rise to a filopodium as a function of their relationship to mitochondria and sites of fission. Same data set as in (E). Fisher’s exact tests.