Fig. 2

Silencing of endogenous Kremen2 expression in NSCLC cells inhibits cell proliferation. A Colony formation assays were conducted using A549-Krm2^KO cells. B Cell proliferation of A549-Krm2^KO and A549-Krm2^KO + Flag-Krm2 was detected by the EdU assay. C Kremen2 expression was stably silenced using shRNA; Kremen2 mRNA expression levels in H1703 and A549 cells were detected by qPCR. D, E The effects of Kremen2 knockdown on colony formation in H1703 and A549 cells. Representative images of the colony formation (D) and clone formation rates (E). F Cell proliferation was measured by the EdU proliferation assay with/without Kremen2 knockdown in H1703 and A549 cells. G A subcutaneous xenograft model of nude mice was established using A549 cells with stable Kremen2 knockdown (n = 12 per group). H, I Tumor volume curve (H) and final tumor weight (I). J IHC analysis of Kremen2 expression on the xenografts (scale bars: 100 μm). The Ki67-positive results (%) are shown in the right panel. Bars represent the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001