Figure 1
Microtubules (MTs) form rings at plus ends and lose connection with nucleus in CENP-F−/− cardiomyocytes. Cardiomyocytes isolated from wild-type (a) and CENP-F−/− (b) mice were fixed and immunostained with DM1A antibody (green) and TOPRO (blue). A zoom in of the circumferentially-oriented MT network (a’,b’) and longitudinal array of MTs (a”) are shown in wild-type and CENP-F−/− mouse cardiomyocytes, respectively. Cardiomyocytes were treated with nocodazole to destabilize microtubules for phenotype comparison (b”). Immunostaining of MTs (green) and nucleus (blue) in wild-type cardiomyocytes show a microtubule-nuclei relationship (c), accompanied by a representative scan (c’). This relationship is completely gone in CENP-F−/− cardiomyocytes (d), accompanied by a representative scan (d’). In (e), quantification of the maximum tubulin and nuclear associated tubulin of wild-type vs. CENP-F−/− cardiomyocytes, show a tight relationship between microtubules and nuclei in wild-type cells opposed to a complete loss of association in CENP-F−/− cells. In wild-type and CENP-F−/− cells, n = 50 or more z-stack images analyzed for maximum tubulin. In wild-type and CENP-F−/−cells, n = 50 or more z-stack images analyzed for nuclear associated tubulin (e). Scale bars: 20 mm. *p < 0.001. Error bars indicate SEM.
