Fig. 4
From: The engineered expression of secreted HSPB5-Fc in CHO cells exhibits cytoprotection in vitro
sHSPB5-Fc decreases the aggregation of GFP-Httex1-Q74 in PC-12 cells. A, Fluorescent image of PC-12 cells that express GFP alone or GFP-Httex1-Q74. The cells were treated with Human IgG (control) or with 50 or 100 μg/ml of sHSPB5-Fc for 24 h. The arrow indicates soluble GFP-Httex1-Q74. The arrowhead indicates insoluble GFP-Httex1-Q74. Scale bars are 50 μm. B, The percentage of cells with GFP-Httex1-Q74 aggregates in A. Three hundred GFP positive cells of each analysis were analyzed. C, Determination of the anti-aggregation properties of sHSPB5-Fc using the membrane-filtration assay. Cell lysates that were treated in same way as in A were immunoblotted using a 0.2 μm cellulose acetate (CA) membrane on a slot bot apparatus. The insoluble GFP-Httex1-Q74 protein on the membrane was immunoblotted with GFP antibody. D, The quantification of protein bands in C versus β-actin were measured with Image J software. Student’s t-test was used for statistical analysis. *p < 0.05, **p < 0.01, ***p < 0.0001