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. 2021 Oct 15;11(10):4788–4806.

Figure 1.

Figure 1

Regulation of DKK1 gene expression in Hep3B and Huh7 cells. A. Effects of DKK1 on invasion assay in HUVECs. NT: no treatment. HUVECs were treated with the indicated concentration of rDKK1 and 50 ng/mL VEGF (as a positive control). B. DKK1 expression levels of human hepatocellular carcinoma cell lines were measured using ELISA (a) and CRISPR-Cas9 based DKK1 knockout was confirmed by PCR (b). C. Western blot analysis of aggregated proteins, collected using centrifugation from Hep3B wild-type, Hep3B CRISPR#73 (#73) and Hep3B CRISPR#131 (#131) cells. DKK1-transfected HEK 293T cells were used as a positive control. D. Protein levels of secreted DKK1 from Hep3B wild type, #73 and #131 cells were measured using ELISA. E. Western blot analysis of aggregated proteins from Hep3B, Huh7, Huh7 Pur-DKK1, Huh7 Pur-Mock cells. Huh7 Pur-DKK1 and Huh7 Pur-Mock cells were cultured in the absence or presence of doxycycline (100 ng/mL) at 37°C for 48 h. Hep3B cells were used as a positive control for DKK1. Lysate: the aggregated fraction after centrifugation; sup: the supernatant (soluble fraction). F. Secreted DKK1 protein levels of CM of Huh7 Pur-Mock and HuH7 Pur-DKK1 cells were measured using ELISA after incubation with doxycycline (100 ng/mL) for 24 h.