Live cell imaging of IPMCs. MDMs were nucleofected to express PH-GFP and imaged after 24 hours using an UltraVIEW Vox spinning disc confocal system, as described in Figure 1 (see Additional files 6 and 7). Images were recorded at 6 frames/minute with a 60× Nikon oil immersion objective (NA 1.4) using Volocity 5.3.2 (Perkin Elmer) and the Perfect Focus System, and analyzed with Volocity, Adobe Photoshop, ImageJ and Fiji software. The white arrowheads in A and B show connections to the cell surface. The images A’, B’ at the right show enlargements of the areas marked by the red boxes. All experiments were repeated on cells from at least three different donors. Scale bars = 20 μm.