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. 2009 Oct 13;4(10):e7438. doi: 10.1371/journal.pone.0007438

Figure 2. Rac inhibition suppresses the pro-fibrotic phenotype of lesional SSc fibroblasts. mRNA and protein analysis.

Figure 2

Fibroblasts were from normal individuals (NF) and individuals with SSc (SScF) were assessed in the presence or absence of the rac-specific inhibitor (NSC23766, 24 hour treatment 50 µM). (A) Real-time PCR analysis. Messenger RNA was harvested from cells and subjected to real-time PCR analysis to detect the mRNAs indicated. Fibroblasts from 6 individuals were analyzed. Data represent averages and standard deviation. As a control, 28S RNA was amplified. Average of three replicates from three separate individuals, adjusted for 28S RNA expression values, are shown (+/− SD * = p<0.05 significant inhibition by NSC23766 compared to untreated controls, Student's paired t test). (B) Western blot analysis. Proteins were harvested and subjected to Western blot analysis with antibodies directed against the proteins indicated. Densitometry is on the right (+/− SD, * = p<0.05 significant inhibition by NSC23766 compared to untreated controls, Student's paired t test) (C) Immunofluorescence analysis. Cells were fixed and stained with anti-vinculin antibody to detect focal adhesion and rhodamine phalloidin to detect actin and α-SMA.