Figure 4
mTORC1 inhibitor rapamycin and ER stressor tunicamycin mobilize different sets of ER-mitochondria connectors.
(A) Immunofluorescence of CNX (green) and MitoTracker Orange staining (mito, red) of control HeLa cells (con) or cells treated with tunicamycin (tun) or rapamycin (rap), measured by confocal fluorescence microscopy. (B) CNX-to-mitochondria colocalization of images acquired as in A, calculated locally as radial Manders’ coefficients (n = 3). (C) Immunofluorescence of PACS2 (green) and MitoTracker Orange staining (mito, red) of control HeLa cells (con) or cells treated with tunicamycin (tun) or rapamycin (rap), measured by confocal fluorescence microscopy. (D) PACS2-to-mitochondria colocalization of images acquired as in (C) calculated locally as radial Manders’ coefficients (n = 3). (E) Immunofluorescence of Mfn2 (green) and MitoTracker Orange staining (mito, red) of control HeLa cells (con) or cells treated with tunicamycin (tun) or rapamycin (rap), measured by confocal fluorescence microscopy. (F) Mfn2-to-mitochondria colocalization of images acquired as in (E) calculated locally as radial Manders’ coefficients (n = 3). Scale bars = 10 μm. Black arrowheads show zones of low colocalization; white arrowheads show zones of high colocalization. Data are shown as the mean ± SEM. *P ≤ 0.05 compared with controls (con).
