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. 2017 Sep 5;7(16):3920–3932. doi: 10.7150/thno.21389

Figure 2.

Figure 2

LSH induces TET2 expression in vitro and in vivo. RT-PCR analysis was conducted to detect TET2 and TET3 genes using total RNA derived from HK1 (A) and HNE3 (B) cells and matching LSH overexpressed cell lines. The level of gene expression was normalized against the housekeeping gene β-actin and is represented as fold change compared with HK1 and HNE2 cells. (C) Two stable LSH knockdown cell lines (shLSH#1 and shLSH#2) were established by transfecting shLSH sequences into C666-1 cells. RT-PCR analysis was used to detect TET2 and TET3 mRNA after knockdown of LSH. The means and s.d. values were derived from three to four independent experiments. A xenograft model of tumor weight was established in nude mice to evaluate the overexpression of LSH in HK1 (D) and HNE3 (E) cells and the control cells. (F) A xenograft model of tumor weight was established in nude mice to evaluate the knockdown of LSH in C666-1 cells. IHC was performed using antibodies against LSH and TET2 in xenograft tissues from HK1 cells (G) and HNE3 cells (H) together with matching LSH ectopic expression of LSH. (I) IHC was analyzed using antibodies against LSH and TET2 in xenograft tissues from C666-1 cells in the depletion of LSH. The mean values of the IHC quantification are shown in the right panel. IHC was performed using antibodies against TET2 in human cancer without metastasis and cancer with metastasis tissues from colon (J) and breast (K). (L) The mean values of the IHC quantification are shown. * p <0.05, ** p <0.01, ***P<0.001.