Fig. 2

PCSK9 promotes B16 melanoma cell-derived oncogenesis. (a) Quantification (mean ± standard deviation/SD) of colony formation by B16 empty vector (EV), PCSK9, D374Y, and Q152H cells based on 3 repeats. See Supplementary Fig. 2b for typical images. Statistical analysis was performed with one-way ANOVA, followed by post-hoc analysis using SPSS. * p < 0.05 in comparison to EV; $$: p < 0.01 in comparison to Q152H. (b, c) Quantification of wound healing and soft agar analyses based on 3 repeats. See Supplementary Fig S2b and 2c for typical images. $ p < 0.05 in comparison to EV; $$: p < 0.05 in comparison to EV, $$$: p < 0.01 in comparison to EV. (d) Quantification (mean ± SD) of Oil Red O staining of the indicated B16 cells. Experiments were repeated 3 time; typical images are presented in Additional file 2: Fig S2f. (e, f) Allografts were produced with the indicated B16 cell lines (5 mice/line) in C57BL/6 mice. Experiments are terminated at the endpoint (tumor volume ≥ 1000 mm³). Tumor volumes (mean ± standard error/SE) (e) and survival curves are prepared (f). Logrank test was performed. *: p < 0.05 and ***: p < 0.001 compared to EV. $$: p < 0.01 and $$$: p < 0.001 in comparison to Q152H; #: p < 0.05 in comparison to PCSK9. (g, h) Oil Red O (ORO) (g) and filipin (h) staining of allografts produced by B16 EV (EV), PCSK9, D374Y, and Q152H cells. Five tumors per genotype were stained; typical images are presented (left panel) along with quantification (right panel). The marked regions were enlarged two-fold. *: p < 0.05, **: p < 0.01, and ****: p < 0.0001