Fig. 4

C/EBPβ enhancer functions to promote HCC tumorigenicity. a The monoallelic (C/EBPβ enh^+/−) and biallelic (C/EBPβ enh^−/−) deletions of C/EBPβ enhancer in HepG2 and LO2 liver cells via CRISPR/Cas9 were confirmed by PCR. b, c qRT-PCR analyses of b C/EBPβ eRNA and c C/EBPβ mRNA levels in WT, C/EBPβ enh^+/− and C/EBPβ enh^−/− cells. C/EBPβ eRNA/mRNA levels were calculated by the 2^−ΔΔCt method using 18s rRNA as internal control, and are presented as fold-changes against the average value of the WT group. d, e Cell proliferation of WT, C/EBPβ enh^+/−, and C/EBPβ enh^−/− cells was determined by colony-formation assays. Representative images of focus formation are shown. f, g Cell invasion of WT, C/EBPβ enh^+/−, and C/EBPβ enh^−/− cells was determined using Matrigel chambers. Representative images of crystal violet-stained invaded cells are shown. h, i Xenograft tumors formed by WT, C/EBPβ enh^+/−, and C/EBPβ enh^−/− cells are shown in the images. Mean xenograft tumor volumes were plotted against days after injection. Data are presented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001 as calculated by unpaired two-tailed Student’s t-test (b–g), and two-way ANOVA (h, i)