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. 2022 Dec 2;15:966209. doi: 10.3389/fnmol.2022.966209

Figure 3.

Figure 3

Miro1 R272Q alters calcium dynamics. (A) Representative Images of hDaNs before and after stimulation with 5 μM Thapsigargin. hDaNs were pretreated for 2 h with 2 μM Mitoxantrone and stained with cytosolic calcium indicator FLUO-4. (B) Cytosolic calcium trace in hDaNs before and after stimulation with 5 μM Thapsigargin. Quantification of total FLUO-4 signal. n = 7–9 (each n corresponds to 1 culture dish) obtained from 3 independent differentiations (nDiff = 3), data displayed as mean ± SEM. (C) Quantification (from Figure 3B) of FMAX/F0, (D) reduction rate of FLUO-4 intensity after reaching peak intensity and (E) the mean baseline FLUO-4 signal. n = 7–9, nDiff = 3, data displayed as mean ± SEM; One-way ANOVA with Tukey’s multiple comparisons for FMAX/F0; Kruskal Wallis with Dunn’s multiple comparisons for the rate of reduction of FLUO-4 signal and mean baseline FLUO-4 intensities. (F) Calcium trace in SH-SY5Y after stimulation with 5 μM Thapsigargin. SH-SY5Ys were transfected with non-targeting or RHOT1 siRNA to knockdown Miro1. SH-SY5Y cells were treated for 2 h with 10 μM Mitoxantrone. Cytosolic calcium was visualized using FLUO-4. Quantification of total corrected signal and normalized relative to t = 0. n = 2–3, data displayed as mean.