Fig. 3.
Myofibroblast activation is Galectin-3 dependent. (a) Galectin-3 expression is up-regulated in myofibroblasts during the hepatic fibrotic response in vivo. Liver sections from WT control (olive oil) (i–iii) and chronic CCL4 injured (8 weeks) (iv–vi) mice were stained with Galectin-3 antibody (green), anti-αSMA (red), and DAPI (blue). (b) Western blot analysis of Galectin-3 expression in primary mouse (mGal-3) and primary human (hGal-3) HSCs during transition from the quiescent to the activated phenotype on tissue culture plastic. (c) Western blot analysis of α-SMA and β-actin expression in WT and Galectin-3−/− primary mouse HSCs cultured on tissue culture plastic for 7 days in 16% FCS. (d) Western blot analysis of α-SMA in WT and Galectin-3−/− primary mouse HSCs after addition of recombinant murine Galectin-3 (30 μg/ml) to Galectin-3−/− HSCs in 16% FCS. (e) Real-time PCR quantitation of procollagen (I) in WT and Galectin-3−/− primary mouse HSCs after addition of recombinant murine Galectin-3 (30 μg/ml) in 16% FCS. ∗, P < 0.05 compared with untreated Galectin-3−/− HSCs. (f) Cell growth of WT (○), Galectin-3−/− (□), and Galectin-3−/− HSCs plus recombinant murine Galectin-3 (30 μg/ml) (■) measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. (g) Internalization of recombinant murine Galectin-3 by Galectin-3−/− primary mouse HSCs. Cells were stained with DAPI (blue) and Galectin-3 antibody (green). (i) Untreated, (ii) 10 min, (iii) 30 min, (iv) 60 min after addition of 30 μg/ml recombinant mouse Galectin-3.