FIG 7.

Miro-1 regulates mitochondrion-dependent calcium influx during adhesion and migration. (A) Flow cytometry analysis of intracellular calcium dynamics (Fluo4-AM staining) in response to the addition of CXCL12 (10 nM) to a suspension of control and Miro-1-silenced CEM T cells (prestained with DiD and DiL, respectively). Results from a representative experiment out of three performed are shown. (B) Representative intracellular calcium oscillation in control and Miro-1-silenced CEM T cells migrating on TNF-α-activated HUVEC. (C) Distribution of relative calcium peak intensity (left) and peak duration in seconds (right). Data are shown as a scatter plot; horizontal lines depict medians from two independent experiments (67 control and 62 Miro-1-silenced cells) (***, P < 0.001). (D) Amounts of ATP in unstimulated and CXCL12-stimulated (10 min) control and Miro-1-intefered cells. Data are means ± SEM from two independent experiments. ***, P < 0.001; *, P < 0.05 (by one-way ANOVA). (E) Time course of the phosphorylation status of ERK1/2 in control and Miro-1-silenced CEM T cells activated with CXCL12. Total protein expression is shown as a loading control. The right panel shows pERK/total ERK band intensity ratios relative to the ratio in control cells at t = 0. Data are presented as the means ± SD from two independent experiments.