Figure 7. The depletion of LRP5, LRP6 or STK40 delays tumor growth.

MDA-MB-468 cells were transfected with control (ctrl, black), LRP5 (^#2, red), LRP6 (^#8, blue) or STK40 (^#5, green) siRNAs. (A) The levels of LRP5 or LRP6 protein were evaluated by western blotting 24 hours after transfection. Actin was used as a loading control. (B) STK40 RNA levels were assessed by RT-qPCR analysis 24 hours after transfection with siRNA (C) Twenty-four hours after transfection, 4×10⁶ MDA-MB-468 cells were injected subcutaneously into Swiss nude mice (7 animals/group). Tumor growth was evaluated twice weekly for one month. The data shown are means + SD. The differences between the control siRNA and the other siRNAs (LRP5, LRP6 or STK40) were tested at each time point using an anova model on the raw data and on the square-rooted data, and adjusted for multiple testing using the Benjamini Hochberg correction (Supplementary Figure 1). Differences were considered significant if the adjusted P value was below 0.05 with both raw and squared-rooted data (indicated by ^*: LRP5 and STK40: days 27 and 30; LRP6: days 20, 23, 27, 30). (D) Tumor volume is indicated for each animal at three time points (8, 20 and 30 days) following the injection of the transfected cells.