Fig. 7.
Subcellular localization of FKBP8 in cardiomyocytes and its interaction with heat shock protein 90 (HSP90).
A, Endogenous FKBP8 in the ER in mouse adult cardiomyocytes. Isolated adult cardiomyocytes from mouse hearts were stained with anti-FKBP8 (green) and anti-KDEL (red) antibodies. Images in boxed areas at higher magnification are shown in right panels. Scale bars, 10 μm and 5 μm, respectively. B, Endogenous FKBP8 in mitochondria in mouse adult cardiomyocytes. The cells were stained with anti-FKBP8 (green) and anti-Tomm20 (red) antibodies. Images in boxed areas at higher magnification are shown in right panels. Scale bars, 10 μm and 5 μm, respectively. C, Endogenous FKBP8 in rat neonatal cardiomyocytes. Cultured rat neonatal cardiomyocytes transfected with tdTomato-ER-3 (red) were stained with MitoTracker (blue), then fixed, and stained with anti-FKBP8 antibody (green). Boxed areas are highlighted in the lower panels. Scale bars are 10 μm and 1 μm. D, Coimmunoprecipitation of FKBP8 and HSP90. Forty-eight h after transfection of HEK293 cells with HA-FKBP8 and Flag-HSP90, cell lysates were immunoprecipitated with anti-HA or anti-Flag antibody in the presence of 1 mmol/L CaCl2 or EGTA. Immunoprecipitates (IP) were analyzed by immunoblotting (IB). E, Coimmunoprecipitation of FKBP8 and HSP90 in mouse heart tissue. Heart homogenates were immunoprecipitated with anti-FKBP8 antibody and subjected to immunoblotting. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)