Fig. 3: Invasion into Caco-2 occurs via a single non-damaging scenario.

A Caco-2 expressing Gal-3 are invaded by a single invasion scenario without Gal-3 recruitment or host cell death. A representative live-cell imaging acquisition of invasions through two successive cells (cell 1 and cell 2) is presented. The six time points correspond to the invasion stages defined in Fig. 1. Phase (single z slice), Gal-3 and CM (maximum intensity projection of seven slices) channels, together with an inset (yellow frame) are presented for each time point. The outlines of the two invaded Caco-2 and their nuclei are highlighted with dashed and dotted yellow lines respectively. The entry point into the Caco-2 layer is marked by an arrowhead. B Three-dimensional representation of CM labelling (magenta) presented in (A) within the entire acquisition volume at a single time point (350 min). Epithelial cell outlines are in orange. A top view (XY) and side view (XZ) are presented together with the volume dimensions. C CM channel insets of two time points from the data presented in (A) are presented together with cross-section (CS) views derived from the dashed lines. D Live-cell imaging of infection of Caco-2 not expressing Gal-3 with a C. albicans strain expressing GFPγ. An average intensity projection of five slices is presented. A local transient ‘inflation’ is observed in the CM channel. An inset (yellow frame) showing a clear separation between the CM labelling and C. albicans when the ‘inflation’ is at its maximum size is presented in XY and in a cross section (CS) derived from the dotted line. As the hypha presented has extended from a previously invaded host cell, it is already entirely surrounded by CM labelling at time 0 min. Scale bars are 10 µm in (A, D), 5 µm in (A) Inset, (C) XY views and (D) inset, and 2 µm in (C) CS views.