Fig. 2: USP22 promotes cell proliferation, migration and invasion, and induces CCA EMT in vitro. | Cell Death & Disease

Fig. 2: USP22 promotes cell proliferation, migration and invasion, and induces CCA EMT in vitro.

From: Operative ubiquitin-specific protease 22 deubiquitination confers a more invasive phenotype to cholangiocarcinoma

Fig. 2

A–B HCCC-USP22, Huh28-USP22, RBE-shUSP22, QBC939-shUSP22 and their controls were cultured and their proliferation was evaluated once a day for 3 consecutive days by CCK8 assay. C–D Each cell line was seeded into six-well plates with 5,000 cells/well. Colonies formed after 14 days were fixed and stained for quantification. E–H Cells were plated in cell inserts with 1 × 104 cells/insert (8 µm pore size) with/without matrigel for invasion or migration assay, respectively. After 24 h, five images per insert were randomly taken and quantified by ImageJ. I–J HCCC-USP22, Huh28-USP22 and their controls were cultured for 48 h. The cells were fixed and immunostained with antibodies to E-cadherin (red) and Vimentin (green), and with DAPI for DNA. Cell lysates from these cell lines were also prepared for immunoblotting analysis with antibodies to E-cadherin, Vimentin, MMP2, MMP9 and GAPDH. K–L RBE-shUSP22, QBC939-shUSP22 and their controls were cultured for 48 h. The cells were fixed and immunostained with antibodies to E-cadherin (red) and Vimentin (green), and with DAPI for DNA. All data are presented as the mean ± SD (*p < 0.05, n = 3).

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