Figure 2. Rat lens explants treated with transforming growth factor beta (TGFβ) exhibit nuclear localization of myocardin-related transcription
factor (MRTF-A). Cells were stained with an Alexa Fluor 568 conjugated MRTF-A secondary (red), fluorescein isothiocyanate
for alpha smooth muscle actin (αSMA; green), and 4', 6-diaminodino-2-phenylindole (DAPI) for the nuclei (blue). The untreated
(control) lens explants exhibited mostly cytoplasmic MRTF-A (A) and no αSMA expression (B). Explants treated with 6 µg/ml TGFβ for 48 h showed predominant nuclear localization of MRTF-A (D) along with robust αSMA expression (E). The corresponding composite picture with DAPI (C, F) demonstrates the difference between nuclear and cytoplasmic localization more effectively. Each scale bar equals 100 μm.
Total protein was extracted from the rat lens culture cells and run on western gels. The western blot data clearly demonstrate
an increase in αSMA expression (red) by the cells after TGFβ treatment (G). Levels of β-tubulin present was used as loading controls. Bands of αSMA were quantified with ImageJ (H). Results show a significant increase *(p<0.05) in αSMA production by the explants following TGFβ treatment (n=6) compared
to the untreated controls (n=4).