Figure 7: The combinational effects of the four miRNAs and their potential targets.

(a) Western blot results showing the expression of PBX3 and α2δ1 in Huh7 cells infected with indicated lentiviruses. (b) The percentage of α2δ1⁺ cells was measured in Huh7 cells infected with indicated lentiviruses using flow cytometry. The data are representative of three experiments. (c) The expression of PBX3 and α2δ1 was assayed in α2δ1⁺ Hep-12 cells overexpressing indicated miRNAs by western blots. (d) Hep-12 cells infected with lentiviruses harbouring indicated miRNAs were assayed for their spheroid formation ability (n=6). Data are the average±s.d. of three independent experiments. *P≤0.05 versus control; #P≤0.05 versus any other group, Student’s t-test. (e) Western blot results showing the expression of PBX3 and α2δ1 in Huh7 cells following indicated miRNA knockdown. (f) The spheroid formation efficiency in Huh7 cells was measured following indicated miRNAs knockdown (n=6). Data are the average±s.d. of three independent experiments. *P≤0.05 versus control; #P≤0.05 versus any other group, Student’s t-test. (g–i) Western blot results showing the expression of indicated proteins in HuH7 cells overexpressing indicated genes. β-actin serves as an internal control. (j) Representative photographs demonstrating the spheroids formed by Huh7 cells overexpressing ADRBK2, PBX3 or both. Scale bar, 250 μm; (k) Huh7 cells overexpressing the indicated genes were measured for their spheroid formation efficiency. Data are the average±s.d. of three independent experiments. P values were calculated by Student’s t-test.