Apotome axio imager z1

For the immunofluorescence analysis of C-terminally Strep-tagged human and mouse MMP-3 and MMP-10 in transfected cells, COS-7 cells (3 x 10 4 ) were plated on coverslips in a 24 well plate and were grown overnight to reach 80% confluence. Fixation and permeabilisation of cells were performed as described previously. 25 COS-7 cells were immunostained using the same antibodies already used for Western blot analysis. After washing twice with PBS/2% BSA, cells were treated for 90 min in PBS/2% BSA at room temperature in the dark with donkey anti-rabbit antibody Cy3 or donkey anti-mouse antibody Cy5 conjugate (Dianova, Hamburg, Germany). Cells were washed twice with PBS, embedded in Fluoro-Gel II (EMS, Hatfield, USA) and images were captured with an Apotome fluorescence imager (ApoTome, Axio Imager Z1, Zeiss, Jena, Germany). Commercial and lab-made antibodies were used as stated in Suppl Tables 1 and2. All antibodies were used at a concentration of 1:200. After washing again with PBS containing 2% BSA, the cells were incubated with the corresponding secondary antibody conjugated with Cy3 or Cy5 fluorophore.