Fig. 3

TG2 and fibronectin are deposited in the ECM by astrocytes. Primary rat astrocytes were plated onto laminin (LAM) coated plastic and were treated for 48 h with a range of inflammatory mediators. TG2 and fibronectin (FN) protein levels in whole cell lysates (WCL) of astrocytes were detected by western blotting (a), which were subsequently semi-quantified (b, c). Each bar represents the mean + standard error of the mean of signal intensities from blots of three separate experiments that were calculated as average percentage compared to control (*P < 0.05). Primary rat astrocytes were treated for 48 h with medium alone as a control condition (CTRL), TGF-β1 or TNF-α+IL-1β, after which astrocytes were removed. TG2 deposition (green) and fibronectin deposition (FN, red) were examined by immunohistochemical staining of the extracellular matrix (d, e, and f). TG2 and fibronectin showed co-localization in the extracellular matrix after TNF-α+IL-1β treatment (indicated by arrows in f). Representative images of three independent experiments are shown. Scale bar represents 50 μm