Fig. 1: Characterization of soraR HCC cells.

A Huh7, B Hep3B, and C HepG2 naive and sorafenib-resistant (soraR) cells were treated with increasing concentrations of sorafenib and subjected to MTT assay after 72 h. The values were expressed as % control considering DMSO-treated samples as 100%. Each assay was performed in triplicate, and each experiment was repeated at least two times. The data represent the mean ± S.D of three independent assays. D Equal amounts of total protein from naive (N) and soraR (SR) HCC cells were analyzed by western blots with the indicated antibodies. E Naive and soraR cells plated in transwell plates were allowed to migrate for 48 h and images were captured using NIS-Elements imaging software in Nikon Eclipse Ti Microscope. Scale bar, 100 µm. For quantitation, cells were counted in four different fields and plotted as bar graphs (F). The data represent the mean ± S.D. of two independent experiments. G Naive and soraR cells were plated in ultra-low attachment plates to form spheres, and the spheres were photographed on day 7 using NIS-Elements imaging software in a Nikon Eclipse Ti microscope. Scale bar, 200 µm. H–K RNA extracted from naive and soraR spheres as described in G were analyzed by qPCR with the indicated genes. Lane 1: Huh7-N, Lane 2: Huh7-SR, Lane 3: Hep3B-N, Lane 4: Hep3B-SR. The data represent the mean ± S.D. of three independent qPCR reactions. Significant differences were determined by t test and indicated as: ns, p > 0.05; *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001.