Gaucher Disease Therapy Figure 3. GCase activity and protein. (A) GCase activity was significantly increased in the treated 4L;C* liver by 3-fold in 20 mg/kg/d or 6-fold in 600 mg/kg/d IFG treatment groups, respectively. In the midbrain GCase was significantly increased by 1.4-fold in 4L;C* mice treated with 600 mg/kg/ d IFG, but not changed with 20 mg/kg/d IFG compared to the untreated group. The GCase activity in the 4L;C* spleen was enhanced by 600 mg/kg/d IFG, whereas both 20 mg and 600 mg/kg/d IFG increased GCase activity in the lung. (B) GCase protein was increased in the liver of IFG (20 or 600 mg/ kg/d) treated 4L;C* mice. GCase protein were determined by immunoblot analyses using anti-mouse GCase antibody and normalized to b-actin signal in the same sample. The levels were presented as percentage relative to GCase in WT mice. All the tissues used in GCase activity and protein analyses were from 28 days old mice. The data represent the mean6S.E. for three mice assayed in triplicate and analyzed by Student’s t-test. ***, p,0.001. doi:10.1371/journal.pone.0019037.g003 IFG suppressed proinflammation Proinflammation is a pathologic response in 4L;C* brain as demonstrated by microglial cell activation and astrogliosis [17]. IFG (20 or 600 mg/kg/d) treated 4L;C* showed decreased CD68 staining of microglial cells at 44 days, indicating suppression of microglial cell activation (Fig. 7A). With the disease progressing to the terminal stages, the CD68 positive microglial cells increased in both dosage groups (Fig. 7A). GFAP signals were increased, indicating astrogliosis. In the 20 mg/kg/ d IFG treated 4L;C* brain stem, GFAP signals were weak at 44 days and increased by 57 days, i.e., terminal stage (Fig. 7B). Immunoblot analyses of GFAP of 44 day midbrain samples showed the reduction of GFAP level in IFG treated mice (Fig. 7C). High dose IFG (600 mg/kg/d) was more effective in PLoS ONE | www.plosone.org 4 April 2011 | Volume 6 | Issue 4 | e19037