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. 2019 Oct 17;13(1):45–60. doi: 10.1007/s12195-019-00602-2

Figure 5.

Figure 5

Ccn4 knockout facilitates B16 cell proliferation at optimal growth conditions. (a) Growth comparison of B16F0 with its knockout cells (left) and B16F10 with its knockout cells (right) during 3-day period in regular 2d culture. Assays were performed in triplicates on 96-well plates and 2000 cells were seeded in each well on day 0 and ended before the cells reached full confluence. (b) Anchorage-independent growth for B16F0 and B16F10 knockout cells. Cells were plated in triplicates in soft agar on 6-well plates. Colonies were fixed and counted after 12 days. (c) 2-day growth comparison of B16F0 and B16F0-KO cells in overconfluent conditions in 2D culture. Assays were performed in triplicates on 96-well plates and 80,000 cells were seeded in each well on day 0. (d) Representative photographs from each group in (c). (e) Cell death comparison after 24 h in Anoikis assay. (f) Cell growth comparison in a total of 72 h in Anoikis assay. (g) Representative photographs from each group in (f) at 72 h. *p value < 0.05; ***p value < 0.001.