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. 2008 Apr 15;29(7):1319–1326. doi: 10.1093/carcin/bgn091

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© The Author 2008. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

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Fig. 2. — Role of GPC3 in HCC cell oncogenecity. (A) GPC3 expression in PLC-PRF-5 cells. Mixed cultures of stable cell line were obtained by transfecting pgpc3-GFP or vector control into PLC-PRF-5 cells. Cell lysates were analyzed by western blot with 1G12. The bracket indicates glycanated GPC3 proteins; the arrow indicates the nascent GPC3 protein and the arrowhead indicates the 65 kDa core protein. (B) Growth rate of the PLC-PRF-5 stable cells. Cells (1 × 10⁴) were seeded in 12-well plates in triplicate and cultured in Dulbecco's modified Eagle's medium without serum. Cells were harvested at 48 h intervals and were counted in triplicates. (C) GPC3 knockdown in HuH-7 cells. Cells were transfected with gpc3 shRNA or control vector (shRNA-EGFP), and 48 h later, cell extracts were analyzed by western blot with 1G12. The bracket indicates glycanated GPC3 proteins, and the arrowhead indicates the 65 kDa core protein. (D) Colony-forming assays for HuH-7 cells. Cells were transfected with either shRNA-EGFP (control) or shRNA-GPC3 cells.