Figure 4
ANXA2 is required for MMc-induced COL6 secretion. (a) HRGECs were transfected with control or ANXA2 siRNA, and 48 h post-transfection, the protein levels of ANXA2 were measured by immunofluorescence (left panel) and Western blotting (right panel). The ratio of ANXA2 intensity of ANXA2 siRNA-treated cells versus control siRNA are shown below the ANXA2 blot. Data were normalized to the intensity of α-Tubulin loading control. (b) HRGECs transfected with control or ANXA2 siRNA were treated with MMc (12 µg/mL, 2 h). After washout, the cells were cultured without MMc for the indicated number of hours, and immunostained with anti-COL6 and anti- γ-H2AX antibodies. (a, b) Scale bar: 50 µm in each panel. (c) The percentage of cells with cytoplasmic COL6 (COL6-positive cells) was measured using the samples of b by fluorescent images. A total of more than 100 cells of four independent fields per experiment was analyzed. Statistical significance was determined by the Mann–Whitney U test. *p < 0.05.
