Primary mouse antibody

Confocal microscopy was used to detect 14-3-3 expression in adult rat cardiomyocytes as described previously 21 (link). Briefly, adult rat cardiomyocytes were isolated and plated on laminin-coated 4-well chamber slides (Lab-Tek). Expression of the 14-3-3 protein was detected using a primary mouse antibody (Santa Cruz, 1:100), and α-sarcomeric actinin was detected using a rabbit antibody (Sigma-Aldrich, 1:100). The secondary antibodies used were Alexa Fluor 594-labeled goat anti-mouse antibody and Alexa Fluor 488-labeled goat anti-rabbit antibody (Invitrogen, 1:500), and the mounting medium contained DAPI (Vector Laboratories). A Carl Zeiss 710 confocal microscope with ZEN software was used for visualizing 14-3-3, α-actinin, and DAPI. Total laser intensity and photomultiplier gain were constant for all the groups and settings, and data were verified by two independent observers who were blinded to the experimental group. A minimum of three coverslips was used for each experimental group, and at least three cell images were acquired from each coverslip.