Figure 3

Overexpression of let-7d in vivo. (A, B) Growth curves and average net weights of xenografts formed by 786O-v (n = 5) and 786O-let-7d cells (n = 5). (C, D) Representative micrographs and quantitative data of metastatic colonies in mice lung. DiI-positive lung metastatic colonies were photographed and counted under a laser confocal microscope. (E) Real-time PCR quantification of relative Alu-sequence expression in mice lung. (F) Quantitative data of the mean CD68 positive cells per five fields in each group. (G) Representative picture of IHC staining of CD68 positive cells. Original magnification: ×200. Data in (A–F) represent the mean ± SD of five mice per group. P values were obtained by the two-tailed Student’s t-test. *P < 0.05. (H) HE staining of the patient tumor and its corresponding derived xenograft (2nd passage). Original magnification: ×200. (I) Real-time RT-PCR analysis of let-7d expression in human RCC tissue, paired normal adjacent tissue, and corresponding PDX tumor tissues 1 week after intratumoral injection of let-7d mimics or control RNA. (J, K) Growth curves and average net weights of xenografts injected with let-7d mimics (n = 7), control RNA (n = 7) or PBS control (n = 7). (L, M) Representative pictures and quantitative data of metastatic colonies in mice lung. (N) Real-time PCR quantification of relative human-specific Alu-sequence expression in mice lung. The values in (J–N) are presented as the mean ± SD of seven mice. P values were obtained by one-way ANOVA. **P < 0.01 (let-7d mimics vs. PBS or control RNA).