Fig. 4
Silencing of AHCYL1 increases the tumorigenic and angiogenic capacities in vivo. A–F NODscid Female mice (n = 8–11 mice per group), and Male (n = 7–8 mice per group) were subcutaneously injected with 2 × 106 A549 cells stably expressing control shRNA (NT) or AHCYL1 shRNA (KD-AL1-4). Average tumor volume ± SD is plotted against time (in days). Different shading corresponds to two independent experiments. Differences were evaluated using a Repeated Measures Design. Final tumor weight and volume are also shown. Means were compared using ANOVA followed by Dunnet’s test. B A549 cells (106, cells, each condition) were intradermal injected in the right flank of NODscid male mice (n = 6–7 mice per group). Vehicle (DMEM without FBS) was injected in the left flank. After 7 days, photographs of skin were taken under magnification glass to quantify vessel density. Representative photograph for each condition (NT vs KD-AL1-4) are shown. Bar: 5 mm. C Quantification (mm2) of vessel density in each condition (n = 13–15 per group). Circles and triangles are used to identify individuals from two independent experiments. Means were compared using a T-test. D Western Blot of VEGF-A (23, 27 and 42 kDa) in NT control and AHCYL1-depleted cells (KD-AL1-4). Quantification of VEGF band (23 kDa) from the western blot of D showing decreased VEGF protein level in AHCYL1 silenced cells (KD-AL1-4). *p < 0.05, **p < 0.01, ***p < 0.001