Alexa fluor 488 or 633 goat anti mouse igg

Immunofluorescence microscopy (IF) was performed using cells adherent on poly L-lysine-coated 24-well glass cover slips. Samples were fixed in 2% paraformaldehyde (pH 7.4) at 37°C for 10 min followed by 3 washes with PBS for 10 min. Slides were blocked with 5% BSA for 1 h at room temperature and stained overnight with mouse anti-Myc at 4°C. Samples were then washed 3 times with PBS, and stained with Alexa Fluor 488 or 633 goat anti-mouse IgG (Invitrogen) or Texas Red Phalloidin (Invitrogen) for 30 min at room temperature. Samples were washed a final 3 times with PBS for 10 min. Slides were mounted with ProLong Gold (Invitrogen) and were imaged on a Leica DM IRE2 inverted fluorescence microscope (Leica, Wetzlar, Germany) and adjusted for brightness and contrast using OpenLab 5.5.0 and ImageJ (NIH). All adjustments were applied equally to all images.