Figure 3.
(A) Microphotographs showing the absence of GS immunostaining in HCC. Surrounding liver (S) shows intense staining of the hepatocytes encompassing the hepatic terminal venules (GS: 5× [left], 10× [right]). (B) qRT-PCR analysis of Glul mRNA levels in tumors (T) compared with the surrounding areas (S). (C) GLS-positive tumors (T) compared with the peritumoral areas (S). (left: 5×, right: 10×). (D) qRT-PCR analysis of Gclc and Glud1 mRNA levels in tumors compared with the surrounding areas. (E) Histochemistry of SDHA showing a remarkable decrease of enzyme activity in c-Myc/h-Ras tumors (outlined areas) compared with the surrounding tissue (left: 5×, right: 10×). (F) Relative SDHA activity. Measurement of SDHA activity was performed using the ImageJ software. (G) complex I activity. The activity of complex 1 was calculated as NADH consumption assessing the difference between the trace slopes with and without complex I inhibitor rotenone (10 μM). Values were then normalized for protein amount. Gene expression is reported as a fold-change of tumor mRNA relative to surrounding livers. Relative quantification analysis for each gene was calculated by the 2ΔΔCt method. The histogram represents means ± SD of 3 tumors and 3 surrounding livers (t test). ∗∗P < .01; ∗∗∗∗P < .0001. ns, not significant.