Figure 2

Stability validation of potential housekeeping markers in protein and gene levels. (A) Western blot demonstrated the expression stability of beta-actin and HSP 60. An equal amount of 25 μg of protein lysate was loaded per lane in order to measure the expression stability of beta-actin and HSP 60 among liver tissues with different conditions. Representative blot images were showed parallel to the histograms with protein intensities analysed in different liver tissues groups. C, Cirrhosis; < 2 cm, small size [less than (<) 2 cm] paired tumour (T) and non-tumour (NT); > 2 cm, large size [greater than (>) 2 cm] paired tumour (T) and non-tumour (NT). Statistical analysis was performed by one-way ANOVA. Data are presented as the mean ± SD. (B) Immunohistochemical staining showed the localizations and the levels of expression of beta-actin and HSP60 in tumourous, T (n = 30); non-tumourous cirrhotic, NT (n = 30); and normal, N (n = 16) liver tissues. Representative photos are shown (magnification: ×400). (C) Real-time qPCR was performed to evaluate the mRNA levels of beta-actin and HSP60 in tumourous, T (n = 20); non-tumourous cirrhotic, NT (n = 20); and normal, N (n = 16) liver tissues. Statistical analysis was performed by one-way ANOVA (p > 0.05). Data are presented as the mean ± SD.