Research Paper Volume 12, Issue 6 pp 5195—5208

Figure 3. Peroxisomal defects in normal fibroblasts expressing progerin. (A) Normal human dermal fibroblast cells were infected by DsRed-LA and DsRed-Pg lentiviruses. The exogenous Lamin A and progerin expression were detected by DsRed fluorescence in the nuclei. Peroxisomes localization was indicated by PMP70 and catalase immunofluorescence staining. Bar = 25μm. (B) Quantification of PMP70 puncta per square unit in fibroblasts overexpressing lamin A and progerin. More than 200 cells from 3 independent experiments were analyzed and the data was represented in Tukey box plot. Boxes show the 25^th, 50^th, and 75^th percentiles and the dots indicate the outliers. (C) Relative fold change of ROS activity measured by DCFDA flow cytometry analysis in fibroblasts overexpressing lamin A and progerin. *, p < 0.05. (D) Quantitative RT-PCR analysis of the relative expression of catalase in fibroblasts overexpressing lamin A and progerin. ***, p < 0.001. (E) Normalized catalase activity in fibroblasts overexpressing lamin A and progerin. n.s., not significant. All experiments were performed using mid-passage cells between p15 to p25. All experiments were repeated at least three times and representative data were shown as indicated.