Figure 5
ATR may link to ANXA2 in MMc-induced COL6 secretion. (a) HRGECs were cultured with DMSO (control) or ATRi for 1 h, then MMc (12 µg/mL) was added in the cultures. Two h after MMc addition, the cells were washed and cultured with or without ATRi for the indicated number of hours. The cells were immunostained with anti-COL6 and anti-ANXA2 antibodies. Scale bar: 50 µm. (b) The ANXA2 signal ratio (nucleus/cytoplasm) were measured for more than 25 cells using the samples of a. (c) HRGECs transfected with control or ANXA2 siRNA were cultured with DMSO (control) or ATRi for 1 h, and then MMc (12 µg/mL) was added in the cultures. Two h after MMc addition, cells were washed, and cultured with or without ATRi for the indicated number of hours. The cells were immunostained with anti-COL6 antibody. The percentage of cells with cytoplasmic COL6 (COL6-positive cells) was measured by fluorescent images. A total of more than 100 cells of four independent fields per experiment was analyzed. (b, c) Statistical significance was determined by the Mann–Whitney U test. **p < 0.01.
