Fig. 1.

SLC4A11 Localizes in the Inner Mitochondrial Membrane. (A) Colocalization of SLC4A11 (green) with Mitotracker Red CMXRos in Human Corneal Endothelial Cells (HCEC). (B) Western blot of mitochondrial and supernatant fractions from HCEC and Mouse Corneal Endothelial Cells (MCEC) and (C) from PS120 fibroblasts transfected with Empty Vector (EV) or hSLC4A11-HA. (D) Transmission Electron Microscopy immunostaining of mitochondria with hSLC4A11-HA 10 nm gold (red arrows) and TOM20 25 nm gold (White arrows) antibodies. (E) Isolated mitochondria from HCEC and (F) PS120-hSLC4A11 were suspended and subjected to different concentrations of digitonin at 4 °C for 30 min, pelleted, supernatant collected and pellet lysed. Western blots of each fraction were probed for HA-tagged SLC4A11, OMM markers TOM20 and VDAC, and IMM markers UCP2 and TIM23 (Supplementary Fig. 1A &B). Release of proteins to the supernatant is plotted as ratio of density of supernatant to total (pellet + sup) band density vs. [Digitonin] (note non-linear scale), n = 3, ±SEM. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)