Fig. 3
M1 macrophage secretome induces stress fiber formation in ASCs. A Representative confocal images of MQ-CM-treated ASCs at 72 h showing increased actin stress fibers (phalloidin) and focal adhesions (vinculin) in MQ-CM- and TGFB1-treated ASCs. Scale bars indicate 20 µm. B Representative immunoblots of MQ-CM-treated ASCs after 72 h analyzing levels of ECM-cytoskeleton signaling enzymes including SMA, phosphorylation of FAK and RGD-recognizing integrins ITGA5 and ITGAV. C Flow cytometry analysis of cell surface expression of ITGA5 and ITGAV (n = 6). D Representative images of a conventional scratch assay showing the migration ability of MQ-CM-treated ASCs. The blue lines indicate the initial scratch lines; the yellow lines indicate the relative migration distance after 24 h. Scale bars indicate 200 µm. Quantification of the relative migration distance was calculated using INCUCYTETM software based on cell confluence within the wound region over time (n = 4). All data are shown as mean ± SEM. Asterisks indicate p values of p<0.05 (*), p<0.01 (**), and p<0.0001 (****). TGFB1 treatment served as a positive control for maximal stress fiber induction