Figure 4.

Proliferation and migration of A549 cells reduced by SHH treatment. (A) Effect of SHH on cell viability of A549 cells cultured with different types of macrophages or treated with molecules secreted from macrophages. Each bar is the mean ± SD obtained from eight independent experiments (n = 8). * p < 0.05 compared to M0 co-culture. ^† p < 0.05 compared to each corresponding control (without SHH). (B) Effect of SHH on wound healing. Phase-contrast images of wound areas at 0 and 24 h after wounding of A549 cells. Bottom panel shows migration ability of A549 cells treated with each macrophage conditioned medium (CM). The CMs were from the culture media of THP-1, M0, M1, M2, and SHH-treated M0 cells. Migrated cells remaining on the bottom surface were stained with 0.5% methylene blue. Scale bars, 500 μm for wound closure and 100 μm for migration. (C) Effect of STAT3 and p38 modulators on wound healing reduced by SHH. The cells were treated with CMs from M1, M2, and SHH-treated M0 cells for 24 h. Scale bar, 100 μm. Bar graphs show percentage of wound closure in the cells treated with the indicated chemicals. Five independent experiments were performed (n = 5). * p < 0.05 compared to each corresponding control. (D) Effect of SHH on cell viability and wound healing increased by M2-CM. SHH were added to A549 cells for 24 h in the presence of M2-CM. Scale bar, 100 μm. Data are shown as the mean ± SD of four independent experiments (n = 4). * p < 0.05 compared to the control.