Figure 7.

miR-148a overexpression or TFR1 knockdown affects anchorage-independent growth of HCC. HepG2 and Huh7 cells were transiently transfected with either a miR-148a mimic, TFR1 siRNAs, TFR1 + miR-148a mimic, Empty + miR-148a mimic or respective negative controls. 24 h post-transfection, cells were mixed with complete growth medium and 0.3% agarose. The cell mixture (10,000 cells/well) was added above the agarose base in 6-well culture plate. After 10 days of incubation, the colonies were imaged under 0.5X magnification using an Olympus SZX12 microscope and counted using ImageJ v.1.51k. Quantitative data are shown in (A–C). Treatments are indicated. Left, representative images of colonies formed; right, quantitation of colonies. Experiments were repeated at least three times. Data represented as mean ± SEM. **P < 0.01, ***P < 0.001, ****P < 0.0001, 2-tailed Student’s t test.