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. 2016 Feb 3;7(9):10228–10242. doi: 10.18632/oncotarget.7169

Figure 7. Matrigel culture remedies the effects of NMU knockdown in RL95-2 cells.

Figure 7

RL95-2 cells without or with NMU knockdown were seeded onto the Matrigel-coated dishes for culture. (A) The growth rates were compared using the AlamarBlue assay. The fluorescence value on day 0 (D0) served as the one-fold control. The results are shown as the mean ± SD. n.s, no significance. (B) Represented images of cell morphology were shown on Day 5 of culture. Scale bar, 100 μm. (C) In the upper panel, the levels of phospho-SRC protein in GFP-knockdown control or NMU-knockdown cells seeded on the standard or Matrigel-coated dishes were compared by Western blotting on Day 5 of culture. Total SRC protein served as the internal control. In the lower panel, the band intensities were further quantified by densitometer in three individual experiments. Fold changes in the phospho-SRC/total SRC level between cells seeded on the standard and Matrigel-coated dishes were normalized and compared. Data are shown as the mean ± SEM. *p < 0.05. (D) The active forms of RAC1, RHOA and CDC42 under the above conditions were pulled down by their affinity beads for Western blotting. The inputs of RAC1, RHOA and CDC42 served as loading controls.