BMAL1-KD inhibits cell migration and invasion of CRC cell lines. (A) A scratch-wound healing assay was applied for cell migration assay. Top, Artificial wounds were made in confluent monolayers of control and BMAL1-KD CRC cell lines. Migration of CRC cells towards the wound, photographed from 24 h to 72 h. The top panel shows one of three independent experiments. Bottom, HCT116 BMAL1-KD and SW480 BMAL1-KD cells displayed lower levels of migratory activity in comparison to control cell lines (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001). No significant differences in migratory activity were observed between BMAL1-KD and control SW620 cells. Data are shown as means ± SEM. (B) Invasion assay. Top, The invasive potential of BMAL1-KD CRC cell lines and their control were analyzed in Matrigel Transwells after 96 h by fluorescence microscopy. DAPI was used to stain the nuclei and to determine the number of invasive cells. A representative image of random fields is shown for each cell type. Scale bar, 50 μm. Bottom, Graphs represent the mean of the number of invaded cells per field in three independent experiments. Only the primary BMAL1-KD CRC cells had a lower invasion capacity than control cells (n = 3, *** p < 0.001). Data are shown as means ± SEM.