Fig. 5

Endogenous TG2 affects the morphological appearance of fibronectin. Fibronectin (FN) aggregates on laminin (LAM) coated plastic were studied after removal of astrocytes that were allowed to produce extracellular matrix during 48 h of TNF-α+IL-1β treatment (a, b). Representative blots of four independent experiments are shown. Each bar represents the mean + standard error of the mean of signal intensities from blots of four separate experiments that were calculated as average percentage compared to control. Primary rat astrocytes were treated with TNF-α+IL-1β for 48 h plated on laminin coated plastic. Immunocytochemical double stainings showed immunoreactivity of fibronectin (FN, red) and TG2 (green) in astrocytes (c, d, e, and f). Astrocytes were co-incubated with TNF-α+IL-1β and the TG2-specific inhibitor ERW1041E (10 μM for 48 h) or DMSO (d) or treated with TNF-α+IL-1β after lentiviral downregulation of TG2 (TG2 knockdown (KD)) compared to scrambled knockdown (scrambled KD) (f). Fibril-like fibronectin deposition (indicated by arrows in c and e) was morphological altered after treatment with ERW1041E and after lentiviral downregulation of TG2. Scale bar represents 20 μm