Fig. 1.

Differential characteristics of untreated LX-2 cells versus those treated with MDI. Light microscopy images from A) LX-2 cells treated with MDI for 72 h and LX-2 cells grown under normal culture conditions (NTC) for 72 h. All imaging was performed at 10X using a light microscope. B) Oil Red O staining of LX-2 cells treated with MDI and cells grown under normal conditions. Yellow arrows depict cells showing representative staining. C) protein levels of alpha smooth muscle actin (ACTA2) in MDI-treated cells and untreated cells (NTC) were detected using western blot analysis. Membranes were incubated with primary antibodies directed against ACTA2 (anti-mouse 1:500 dilution; Invitrogen) or GAPDH (anti-rabbit 1:1000; Cell Signaling Technology), washed in 1X TBS-Tween buffer and then incubated with secondary antibodies conjugated to horseradish peroxidase (1:3000; Cell Signaling Technology) for 1 h at room temperature. Membranes were incubated with Clarity Western ECL Blotting Substrate (Bio-Rad; Hercules, CA) according to the manufacturer's instructions. Protein bands were imaged using the Odyssey FC imaging system (LI-COR Biotechnology; Lincoln NE).