Fig. 5. Quantification of gap junction coupling by intercellular small molecule diffusion. To demonstrate gap junction coupling, individual myofibroblasts (A) and fibroblasts (B) in confluent monolayer were microinjected with Lucifer Yellow (yellow, *) and stained after diffusion for 5 minutes for cell nuclei (blue). Scale bar: 25 µm. (C) To quantify diffusion of small molecules through gap junctions, monolayers of myofibroblasts and fibroblasts were scrape-loaded with Lucifer Yellow (green) and neurobiotin (red) in control conditions (n=34) and in the presence of gap junction uncoupling agents palmitoleic acid (PA; n=11) and carbenoxolone (CBX; n=11). Scale bar: 100 µm. The number of cell layers receiving Lucifer Yellow (D) and neurobiotin (E) from the scrape wound was quantified; shown are means ±s.d. per experiment. *P0.001 compared with the respective control.