FIGURE 4.

The expression of aconitase 1 and the total aconitase activity negatively correlate with the pro-SPC expression in IPF (A) Western blot analysis of pro-SPC using whole lysates prepared from the native lungs of the recipients of lung transplantation performed against IPF. Each lane represents an individual sample collected from different sampling sites (please see Figure 3A for the sampling method). Equal amounts of protein (7.5 μg) were loaded per lane. A common reference sample from Patient No.3 was loaded at the last lane for each blot to enable the quantitative comparison between different blots. LU (left upper lobe); LL (left lower lobe) (B) Scatterplot of correlation between aconitase activity and the expression of 12-kD processing intermediate of pro-SPC (#2 in (A) in lung samples collected from IPF patients (n = 18). The correlation coefficient (r) and p-value (p) are shown. *p < 0.05. A p-value of 0.05 or lower was considered to be statistically significant (C) Relative expressions of 12-kDa processing intermediates of pro-SPC (#2 in (A) [upper lobe samples from IPF (n = 6) vs. lower lobe samples from IPF (n = 12)] were shown as a box-whisker plot. Each box shows the upper and lower quartile with the central bar representing the median and the whiskers showing the minimum and maximum (D) Specificity of the pro-SPC antibody used in this article (cat# AB3786, SigmaMillipore) was evaluated by Western blot analysis using whole cell lysates from A549 cells transfected with either pro-SPC-overexpression or control plasmids. Equal amounts of protein (7.5 μg) were loaded per lane. Data from one representative experiment of two or more independent experiments are shown.