Figure 6.
(A) Representative staining patterns in human HCC specimens for LDHA. In normal liver (NL), a faint or absent staining for LDHA was detected. Surrounding nontumorous liver tissues (ST) exhibited weak to moderate cytoplasmic LDHA immunoreactivity, which was generally lower than in corresponding HCC lesions. In this case (HCC1), a steatotic HCC shows a more pronounced LDHA immunolabeling than the neighboring ST. HCC2 depicts instead a well-differentiated HCC displaying weak immunoreactivity for LDHA. Tumors with moderate (HCC3) and poor (HCC4) differentiation were instead most frequently characterized by robust LDHA immunoreactivity. Strong immunolabeling for LDHA was often observed not only in the cytoplasm but also in the membrane (HCC3) and the nucleus (HCC4). Original magnification = 200×. scale bar = 100 μM. (B) Representative Western blot analysis results for LDHA protein levels in human HCC. The blot of a series of 7 tumors (T) and corresponding nontumorous surrounding livers (NT), whose quantifications are shown in panel B, is depicted here. Specifically, tumors 1–4 and 7 displayed higher LDHA levels than corresponding nontumorous livers, whereas tumors 5 and 6 exhibited lower levels than respective non-neoplastic livers. The lower panel shows equal loading in the various samples as assessed by reversible Ponceau red staining. (C) Graph showing the results of Western Blot analysis of the levels of LDHA protein in human HCCs and their respective peritumoral counterparts. (D) qRT-PCR analysis of LDHA. qRT-PCR analysis was performed on the same samples shown in B. Relative quantification analysis for each gene was calculated by the 2ΔΔCt method. M, marker