Figure 3.

Rh2 inhibited SASPs-induced migration and invasion of breast cell lines. (A) MDA-MB-231 cells were cultured with conditioned medium from senescent MDA-MB-231 and MCF-10A cells treated with or without Rh2 for 48 h. The cell-free areas were imaged with microscope at 0 h and 48 h, respectively. The changes in cell-free area were calculated using CellProfiler Software (2.2.0). At least three wound scratches were analyzed per experiment. (B) Invasion of MDA-MB-231 cells (5 × 10⁴/well) as determined by 24-well plate transwell system. Cells on the lower side of membranes were stained and quantified. (C) Western blot analysis of representative epithelial–mesenchymal transition (EMT) markers of MDA-MB-231 cells after CM treatment. Immunostaining for the tight junction protein ZO-1(Red) and the nuclear regions were counterstained with DAPI (blue). * indicates p < 0.05 versus Nontreated con, ** indicates p < 0.01 versus Non-treated con; *** indicates p < 0.001 versus nontreated con. ^## indicates p < 0.01 versus CM alone group. ^### indicates p < 0.001 versus CM alone group.