Figure 5.

Localisation of calcein in CMs and CFs and vimentin immunofluorescence in co-cultures. (A) Freshly isolated CMs were loaded with 5 μM calcein for 30 min at 37 °C and washed three times in PBS. 1×10⁴ rod shaped CMs in 0.5% FBS media were then added to a CF monolayer in 35 mm glass bottomed petri dishes. The localisation of calcein was visualised immediately following CM addition (0–1 h) then again following 20 h (upper and middle panels respectively). The areas surrounded in the red box in the middle panel are expanded in the bottom panel showing calcein dye transfer to the underlying CFs depicted by white arrows. Red arrows show examples of areas of the CM being pulled by the underlying CFs. Images were taken on the Zeiss 510 LSM confocal microscope. Scale bar represents 100 μm on the upper and middle panels and 50 μm on the lower panel (n=3). (B) Cultures were fixed following 24 h following of co-culture and stained for vimentin. All images were taken at x25 magnification on the Zeiss 510 LSM confocal microscope and normalised to an IgG control. Scale bar represents 100 μm (n=3).