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Fig. 2 | Molecular Cancer

Fig. 2

From: Lipocalin2 suppresses metastasis of colorectal cancer by attenuating NF-κB-dependent activation of snail and epithelial mesenchymal transition

Fig. 2

Negative effects of LCN2 on proliferation and tumorigenicity. a LCN2 expression confirmed by western blots and real-time PCR in the indicated cell. (s: supernatant) and quantification of relative mRNA expression of LCN2 expression of corresponding cells. b Cell proliferation measured by the CCK8 assay, and quantification of OD value at 450 nm of corresponding cells with microplate reader (5000 cells/well, mean ± SD from at least three independent experiments). (C,D) Colony-formation assays on soft agar (c) and plate (d) with corresponding cells and quantification of the fold change of average clone diameter of eight fields (for soft agar colony-formation assay) and relative counts based on the colony diameter (for plate colony-formation assay, SW480-SHB/sh-LCN2: diameter 5 cm; SW620-OB/LCN2: diameter 2 cm) (mean ± SD from at least three independent experiments. Scale bars, 100 μm). e Tumor weights (g) and volumes (mm3) of mouse xenografts formed with corresponding cells. SW620-OB (5 × 106), SW620-LCN2 (5 × 106), SW480-SHB (1 × 107), and SW480-sh-LCN2 (1 × 107) cells in 100 μl PBS were inoculated subcutaneously into the left shoulders of 4 to 5-week-old BALB/c nude mice. Tumor length and width were measured with calipers, and tumor volumes were calculated according to the equation: V (mm3) = ½ [width2 (mm2) × length (mm)]. Growth curves were plotted based on mean tumor volume within each experimental group at the indicated time points. Tumor growth was observed for 27 days (for SW620-OB and SW620-LCN2 cells) and 16 days (for SW480-SHB and SW480-sh-LCN2 cells), respectively. * P < .05, ** P < .01, *** P < .001

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