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. 2019 Sep 23;9:13703. doi: 10.1038/s41598-019-50275-0

Figure 6.

Figure 6

Phosphorylation of ADD1 at T724 is important for cell migration and cell invasion. (a) MDA-MB-231 cells were infected with lentiviruses expressing shRNAs specific to ADD1 (sh-ADD1) or luciferase (shLuc). FLAG-ADD1 or its mutants were re-expressed in the ADD1-depleted cells (sh-ADD1/FLAG-ADD1). The cells were suspended in serum-free medium and subjected to the cell migration assay with (+) or without (−) 40 ng/ml EGF in the lower chambers. After 6 h, the migrated cells were fixed, stained, and counted using a light microscope. Representative micrographs are shown. Bars, 100 μm. The number of migrated cells were measured and expressed as percentage relative to the control in the absence of EGF. Values (means ± s.d.) are from three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001. (b) MDA-MB-231 cells described in (a) were suspended in serum-free medium and subjected to the invasion assay with (+) or without (−) 40 ng/ml EGF in the lower chambers. After 24 h, the cells that migrated through Matrigel were fixed, stained and counted using a light microscope. Representative micrographs are shown. Bars, 100 μm. The number of cells were measured and expressed as percentage relative to the control in the absence of EGF. Values (means ± s.d.) are from three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.