Skip to main content
. 2020 Sep 23;12(10):2734. doi: 10.3390/cancers12102734

Figure 3.

Figure 3

Involvement of HDAC9 in the sphere formation capacity of undifferentiated HCC cells. (a) Cell proliferation assays of HLE (left), HLF (center), and Hc (right) cells treated with si-Control (open circle) and si-HDAC9 (closed circle) for 3 days. Cell viability was determined by water-soluble tetrazolium (WST) assay and expressed as the relative amount of viable cells compared to day 0. * p < 0.05, ** p < 0.01 vs. si-Control group, N.S.: not significant, n = 3 (b) Morphological appearance of spheres formed in the culture medium containing 10 pmol/mL si-Control (left) and si-HDAC9 (right). HLE (upper) and HLF (lower) cells were cultured in an ultra-low attachment culture plate for 7 days and photographed. Scale bar = 200 µm. (c) The diameter of the sphere was measured in ten representative spheres of each group. ** p < 0.01 vs. si-control. (d) Relative gene expression levels of HDAC1, HDAC2, HDAC3, HDAC8, and HDAC9 compared to a house keeping gene (β-actin) were measured in 2D-(monolayer) and 3D-(sphere) cultured HLE and HLF cells by quantitative PCR analysis (n = 4). * p < 0.05, ** p < 0.01 vs. 2D-cultured group. Quantitative PCR analysis was performed at n = 4.