Figure 1 of Xing, Mol Vis 2009; 15:1827-1834.


Figure 1. Hypoxia suppresses TGFβ-induced myofibroblast transformation. A: Primary cultured rabbit corneal keratocytes on coverslips were treated with hypoxia for 4 h each day for 72 h with or without TGFβ1. After treatment, cells were stained for α-SM actin (green) and nuclei (DAPI, blue). Microscope images are representatives of indicated groups (magnification: 200×). Five randomly selected fields were taken from each coverslip. The experiment was repeated three times. The bar graph shows the percentage of α-SM actin positive cells over total cell count in each group from immunofluorescence analysis. Error bars represent the standard error of the mean (n=3 experiments). The asterisk indicates that the indicated groups were significantly different from TGFβ (p<0.05). B: Primary rabbit keratocytes on 100 mm Petri dishes were treated the same as in (A), stained for α-SM actin and then analyzed by flow cytometry. Representative images show α-SM actin fluorescence on the x-axis and forward scatter on the y-axis. Bar graph shows the percentage of α-SM actin positive cells over total cell count in each group. The experiment was repeated three times. Error bars represent the standard error of the mean (n=3). The asterisk denotes that the indicated groups were significantly different from TGFβ (p<0.05). C: Primary rabbit keratocytes on the Petri dish were treated the same as in (A). Whole cell lysates were collected immediately after treatment and analyzed by western blot for α-SM actin. The image shown is the representative western blot of three experiments.