Figure 1
From: Nestin depletion induces melanoma matrix metalloproteinases and invasion
Nestin affects melanoma MMP expression. (a) Nestin mRNA levels were significantly downregulated in nestin KD cells (NesKD, black bars) of A2058 (89.5% decrease) and A375 (91.4% decrease), as compared with vector control cells (Vec, white bars). (b) Decreased nestin protein levels were confirmed in nestin KD A2058 (92.7% decrease) and A375 (97.7% decrease) cells as compared with vector control cells by western blot. (c) Nestin KD did not significantly alter cell proliferation in A2058 and A375 cells. (d) Compared with A2058 vector control cells (white bars), A2058 nestin KD cells (black bars) had significantly higher mRNA levels of MMP1, MMP3, MMP9, and MMP10. (e) Compared with A375 vector control cells (white bars), A375 nestin KD cells (black bars) had significantly higher mRNA levels of MMP1, MMP3, MMP9, and MMP10. (f) Western blot confirmed that A2058 nestin KD cells had marked higher protein levels of MMP1 (3.2-fold) and MMP3 (31.2-fold) in cell lysate and secreted MMP1 (3.0-fold), MMP3 (19.9-fold), and MMP10 (60.7-fold) in supernatant. MMP9 protein in supernatant of A2058 cells was below detection limit of immunoblotting. A375 nestin KD cells had marked higher protein levels of MMP1 (7.3-fold) in cell lysate and secreted MMP1 (4.4-fold) and MMP9 (11.1-fold) in supernatant. MMP3 protein in cell lysate and supernatant of A375 cells were below detection limit of immunoblotting. MMP9 and MMP10 proteins in cell lysate of A2058 and A375 cells were below detection limits of immunoblotting and thus data were not shown.
