Figure 7: Gαi1 and Gγ2 proteins have limited effects on Gβ2-induced mitochondrial aggregation.
From: G-protein β2 subunit interacts with mitofusin 1 to regulate mitochondrial fusion
(a) Upper panels: Gβ2 shRNA HeLa cells (red) were transfected with GFP-Gαi1 (green) and loaded with 20 nM MitoTracker Deep Red; bottom panels: HeLa cells were co-transfected with GFP-Gαi1 and Flag-Gβ2 (red) and loaded with 20 nM MitoTracker Deep Red. (b) HeLa cells were transfected with Flag-Gγ2, Gβ2-c-myc and YFP-Gαi1. After incubation for 24 h, the cells were incubated with Mitotracker (Deep Red), fixed and immunostained with Flag antibody (red) and c-myc antibody (green). (c) The histogram indicates the mitochondrial aggregation percentage in the cells transfected with either of the coupled pairs: Flag-Gβ2 alone, Flag-Gβ2 and Gγ2, Flag-Gβ2, Gγ2 and YFP-Gαi1, Flag-Gβ2 and YFP-Gαi1, YFP-Gαi1 alone and Gγ2 alone, respectively. Overall, 350–400 cells were counted in each experiment. Error bars represent the s.e.m. (d) HeLa cells were transfected with Gγ2 and Flag-Gβ2 or Flag-Gβ2 (Δ1–40). After incubation for 24 h, the cells were incubated with Mitotracker (Deep Red), fixed and immunostained with Flag antibody and Gγ2 antibody. (e) Gβ2 shRNA HeLa cells (red) were transfected with Flag-Gγ2 (green) and loaded with 20 nM MitoTracker Deep Red. The scale bar in all images represents 10 μm. (f) The histogram indicates the mitochondrial aggregation percentage in the cells transfected with either of the coupled pairs: Flag-Gβ2 alone (open box), Flag-Gβ2 (Δ1–40) (grey box), Flag-Gβ2 and Gγ2 (open box), Flag-Gβ2 (Δ1–40) and Gγ2 (grey box), Gγ2 alone and null vector (filled box), respectively. Error bars represent the s.e.m. Quantification was carried out as described in c.
