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. 2015 Jun 4;6(25):21421–21427. doi: 10.18632/oncotarget.4070

Figure 3. ANX A3 silencing inhibits migration of MCF7 cells.

Figure 3

MCF7 cells were transfected with 10 nM ANX A3-siRNA using Oligofectamine™ (Invitrogen) as per manufacturer's instructions. a. At indicated time points cell lysates were assessed by immunoblotting for ANX A3 and GAPDH as a loading control. The signal was analysed using a LICOR Odyssey imaging system. ANX3 was successfully silenced for the duration of the experiment. b. ANX A3 was silenced in MCF7 cells and cells were seeded onto the upper layer of the transwell cell permeable membrane transwell. At indicated time points cells were fixed, cells on the upper layer of the transwell membrane were removed whereas cells that had migrated to the underside of the transwell membrane were stained with DAPI stain and counted. ANX A3 visibly reduced the number of cells that migrated to the underside of the transwell membrane. c. Quantification of migration showed that migration was significantly reduced in ANX A3-silenced cells at both 48 h (*p < 0.01, n = 3) and 72 h (*P < 0.001, n = 3). d. 5000 cells were seeded into wells of 24 well plates, fixed at indicated time points and stained with DAPI central fields of view were images and cells counted. No difference in proliferation was observed between ANX A3silenced cells and control cells treated with ntg-siRNA (n = 3).